R/ChipSeq.R
In aiminy/ChipSeq: An R Package for Performing ChipSeq Analysis In One Step
#' AnnotatePeak2
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#' input.file.pattern="*macs142_peaks.bed"
#' output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#'
#' AnnotatePeak(input.file.dir,input.file.pattern,8,output.file.dir,genome="Mm")
#' AnnotatePeak(input.file.dir,input.file.pattern,7,output.file.dir,genome="Hs")
#'
#'
#'
AnnotatePeak2 <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",Sys.time())
#temp2=gsub(" ","-",temp1)
#temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
temp3=output.dir.name
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
#' AnnotatePeak3
#'
#' @param input.file.dir
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#' output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#'
#' AnnotatePeak3(input.file.dir,output.file.dir,genome="Mm")
#' AnnotatePeak3(input.file.dir,output.file.dir,genome="Hs")
#'
#'
#'
AnnotatePeak3<- function(input.file.dir,output.file.dir,genome) {
library(ChIPpeakAnno)
re<-ParserReadFiles(input.file.dir,"bed",output.file.dir)
#print(file.name.2)
file.name.2<-re$input
output.dir.name=output.file.dir
#dir.name=input.file.dir
#input.file.pattern=input.file.pattern
#dir.name=reformatPath(dir.name)
#output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",Sys.time())
#temp2=gsub(" ","-",temp1)
#temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
temp3=output.dir.name
dir.create(temp3)
#file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
#file.name.2<-as.list(file.name)
#names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
#print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
#' AnntationUsingChipSeeker
#'
#' Use ChipSeeker to annotate data
#'
#' @param dir.name: path for bed files
#' @param input.file.pattern : input file pattern
#' @param out.dir.name output file directory
#' @param txdb Annotation databased to be used("hg19","hg38")
#' @param DD distance definition around TSS
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed"
#' input.file.pattern="common_peaks.bed"
#' out.dir.name="/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnntationUsingChipSeeker <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=5000,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
#re<-ParserReadFiles(dir.name,input.file.pattern)
#re.bed<-re$input
#re.peaks.only.bed.2 <- re.bed
file.1 <- list.files(dir.name,pattern=input.file.pattern,all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
#re.bed<-re$input
re.peaks.only.bed.2 <- file.1
# if(length(dir(dir.name,pattern="peaks.bed"))!=0)
# {
# re.peaks.only.bed.2<-FL(re.bed,'peaks')
# cat("peaks\n")
# print(re.peaks.only.bed.2)
# }
#
# if(length(dir(dir.name,pattern="summits.bed"))!=0){
# re.summits.only.bed<-FL(re.bed,'summits')
# cat("summits\n")
# print(re.summits.only.bed)
# }
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,"Annotation",APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
res <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
#select.index <- which(peakAnno$distanceToTSS<20,000 && peakAnno$distanceToTSS >= -20,000)
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
x_name=names(re.peaks.only.bed.2)[u]
cat(x_name)
png(file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_pie.png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
#print(paste0(peaks[,c(2,3)]))
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_only_mapped_peaks.xls")),
row.names = FALSE,quote=FALSE,sep="\t")
unmapped.peaks<-peaks[-which(paste0(peaks[,2],"_",peaks[,3]) %in% paste0(peaks.anno[,2],"_",peaks.anno[,3])),]
cat(dim(peaks)[1]," ",dim(peaks.anno)[1]," ",dim(unmapped.peaks)[1],"\n")
if(dim(unmapped.peaks)[1]!=0){
colnames(unmapped.peaks)=colnames(peaks.anno)[1:6]
unmapped.peaks.3<-smartbind(peaks.anno,unmapped.peaks)
unmapped.peaks.4<-unmapped.peaks.3[order(unmapped.peaks.3[,1],unmapped.peaks.3[,2]),]
write.table(unmapped.peaks.4,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_all_peaks.xls")),row.names = FALSE,quote=FALSE,sep="\t")
}
re <- list(peaksMappedOnly=peaks.anno,peaksAll=unmapped.peaks.4)
re
},re.peaks.only.bed.2,d)
res
}
#' BamFileSortIndexVisualization
#'
#' @param input.file.dir
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' output.file.dir="/scratch/projects/bbc/aiminy_project/"
#' genome="Hs"
#' d=4000
#' BamFileSortIndexVisualization(input.file.dir,output.file.dir,4000,genome)
#'
#'bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::BamFileSortIndexVisualization("/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA","/scratch/projects/bbc/aiminy_project/7_27_2017",400000,"Hs")'
#'
#'
BamFileSortIndexVisualization <- function(input.file.dir,input.pattern,output.file.dir,d,genome,bam.sort=FALSE) {
#library(ChIPpeakAnno)
#re<-ParserReadFiles(input.file.dir,"bam")
file.1 <- list.files(input.file.dir,pattern=input.pattern, all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
file.name.2<-file.1
#output.dir.name=re$output
#temp3=paste0(output.dir.name,"_visualization")
temp3=output.file.dir
if (!dir.exists(temp3))
{
dir.create(temp3, recursive = TRUE)
}
#dir.create(temp3)
re.out<-file.name.2
if(bam.sort==FALSE){
cmd1="samtools sort"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd2=paste0(cmd1," ",x," ","-o"," ",file.path(temp3,paste0(x_name,"_sorted.bam")))
print(cmd2)
system(cmd2)
},re.out,temp3)
cmd3="samtools index"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd4=paste0(cmd3," ",file.path(temp3,paste0(x_name,"_sorted.bam")))
#print(cmd2)
system(cmd4)
},re.out,temp3)
}
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
#cmd7="-L 4000"
cmd7=paste("-L",d,sep=" ")
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd8=paste(cmd5,file.path(temp3,paste0(x_name,"_sorted.bam")),cmd6,file.path(temp3,x_name),cmd7,sep=" ")
print(cmd8)
system(cmd8, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
}
convertBam2StrandBw2 <- function(input.bam.file.dir, output.bw.file.dir, BigMem = FALSE,
cores = 15, Memory = 25000, Wall.time = "72:00", span.ptile = 8)
{
re <- parserreadfiles(input.bam.file.dir, "bam")
res <- re$input
m.id <- grep("login", system("hostname", intern = TRUE))
if (!dir.exists(output.bw.file.dir))
{
dir.create(output.bw.file.dir, recursive = TRUE)
}
# job.name=paste0('bamSort[',length(res),']')
cmd.l <- lapply(1:length(res), function(u, m.id, Wall.time, cores, Memory,
span.ptile, res, output.bw.file.dir)
{
# path_name = dirname(res[[u]]) path_name2 <- basename(path_name)
file_name = file_path_sans_ext(basename(res[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
if (BigMem == TRUE)
{
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
} else
{
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' aimin.yan@med.miami.edu",
sep = " ")
}
job.name = paste0("bam2wig.", u)
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.",
job.name, ".log "), paste0("-e %J.", job.name, ".err -W"))
# job.name=paste0('bamSort[',length(res),']') cmd1 = paste0('bsub -w
# \'done(\'bamSort[*]\')\'', 'bsub -P bbc -J \'',job.name,paste0('\'
# -o %J.log '),paste0('-e %J.err -W')) job.name=paste0('Bdg[',u,']') cmd1 =
# paste0('bsub -w \'done(\'bamIndex[*]\') && done(\'Chrosome\')\'',
# 'bsub -P bbc -J \'',job.name,paste0('\' -o %J.',job.name,'.log
# '),paste0('-e %J.',job.name,'.err -W'))
if (u <= 6)
{
cmd2 = paste("bam2wig.pl -pe --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
} else
{
cmd2 = paste("bam2wig.pl --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
}
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
if (u <= 6)
{
cmd3 = paste("bam2wig.pl -pe --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
} else
{
cmd3 = paste("bam2wig.pl --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
}
}
cmd <- cmd3
system(cmd)
cmd
}, m.id, Wall.time, cores, Memory, span.ptile, res, output.bw.file.dir)
}
#'R -e 'library(ChipSeq);ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot")'
#'R -e 'library(ChipSeq); x <- ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot",cores = 8, Memory = 16000,span.ptile = 4,wait = FALSE)'
#'R -e 'library(ChipSeq); x <- ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot",job.option = "parallel", cores = 32, Memory = 16000,span.ptile = 16,wait = TRUE)'
plotBam <- function(input.file.dir,file.type,output.file.dir,job.option = "general",cores = 15, Memory = 25000, Wall.time = "72:00", span.ptile = 8,wait=TRUE) {
#library(ChIPpeakAnno)
#cat(job.option,"\n")
#job.option <- as.character(job.option)
#cat(job.option,"\n")
re<-ParserReadFiles(input.file.dir,file.type)
file.name.2<-re$input
#output.dir.name=re$output
re.out<-file.name.2
m.id <- grep("login", system("hostname", intern = TRUE))
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp3= output.file.dir
#cmd1="samtools sort"
cmd.1 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamSort.", u)
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.",
job.name, ".log "), paste0("-e %J.", job.name, ".err -W"))
cmd2=paste("samtools sort",re.out[[u]],file.path(temp3,paste0(file_name, "_sorted")),sep=" ")
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
cmd3 = paste("samtools sort",re.out[[u]],file.path(temp3,paste0(file_name, "_sorted")),sep=" ")
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
cmd.2 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamIndex.", u)
wait.job.name = paste0("bamSort.", u)
if(wait == TRUE){
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
}else{
cmd1 = paste0("bsub -P bbc -J \"",job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
}
cmd2=paste("samtools index",file.path(temp3,paste0(file_name, "_sorted.bam")),sep=" ")
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
cmd3 = paste("samtools index",file.path(temp3,paste0(file_name, "_sorted.bam")),sep=" ")
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
cmd.3 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamPlot.", u)
wait.job.name = paste0("bamIndex.", u)
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
cmd8=paste(cmd5,file.path(temp3,paste0(file_name,"_sorted.bam")),cmd6,file.path(temp3,paste0(file_name,"_sorted")),cmd7,sep=" ")
cmd3 = paste(cmd1, cmd0, cmd8, sep = " ")
} else
{
"ngs.plot.r -G hg19 -R tss -C config.txt -O 2017-03-02-01_S5_R1.tss -T 2017-03-02-01_S5_R1 -L 4000 -RR 1"
"ngs.plot.r -G hg19 -R tss -C config.txt -O 2017-03-02-01_S5_R1.tss -T 2017-03-02-01_S5_R1 -L 4000 -RR 1 -GO max -SC global"
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
cmd8=paste(cmd5,file.path(temp3,paste0(file_name,"_sorted.bam")),cmd6,file.path(temp3,paste0(file_name,"_sorted")),cmd7,sep=" ")
cmd3 = cmd8
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
#re <- list(cmd.1 = cmd.1,cmd.2=cmd.2,cmd.3=cmd.3)
# return(re)
}
#' DrawVenn
#'
#' @param devtools
#' @param install_github
#' @param Vennerable
#' @param venneuler
#' @param VennDiagram
#'
#' @return
#' @export
#'
#' @examples
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#' DrawVenn(out.dir.name)
#'
DrawVenn <- function(out.dir.name) {
devtools::install_github("js229/Vennerable");
library(Vennerable)
BB=c(1:6153,6154:25736)
CC=c(6154:25736,25737:29579)
AA=c(6026:6153,6154:8462,25737:25776,29650:29719)
Vstem <- Venn(list(ASXL1=AA,SMC1A=BB,RAD21=CC))
SetLabels <- VennGetSetLabels(Vstem)
Cstem3 <- compute.Venn(Vstem,doWeights=TRUE)
SetLabels <- VennGetSetLabels(Cstem3)
FaceLabels <- VennGetFaceLabels(Cstem3)
FaceLabels[FaceLabels$FaceName=="010","x"] <- 10
FaceLabels[FaceLabels$FaceName=="010","y"] <- 3
SetLabels[SetLabels$Label=="SMC1A","hjust"] <- "right"
SetLabels[SetLabels$Label=="SMC1A","x"] <- 110
SetLabels[SetLabels$Label=="SMC1A","y"] <- 58.78653
SetLabels[SetLabels$Label=="RAD21","x"] <- -80
SetLabels[SetLabels$Label=="RAD21","y"] <- 58.78653
SetLabels[SetLabels$Label=="ASXL1","y"] <-80
Cstem3 <- VennSetSetLabels(Cstem3,SetLabels)
Cstem3<-VennSetFaceLabels(Cstem3,FaceLabels)
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="010"),]$x<-90
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="010"),]$y<--35
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="001"),]$x<--90
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="001"),]$y<--35
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$x<--18
#Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$y<--
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$vjust<-"center"
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="110"),]$x<-19
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="110"),]$y<-49.86767
c.3.set=sum(Cstem3@IndicatorWeight[,4])
Cstem3@IndicatorWeight[which(row.names(Cstem3@IndicatorWeight)=="000"),4]=35000-c.3.set
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="DarkMatter"),]$y<--130
temp<-ol$venn_cnt
index.A=grep(colnames(Cstem3@IndicatorWeight)[1],colnames(temp))
index.B=grep("SMC1",colnames(temp))
index.C=grep("Rad21",colnames(temp))
temp2<-as.data.frame(temp[,c(index.A,index.B,index.C,4)])
colnames(temp2)[1:3]=colnames(Cstem3@IndicatorWeight)[1:3]
row.names(temp2)<-with(temp2,paste0(temp2$ASXL1,temp2$SMC1A,temp2$RAD21))
row.names(temp2)<-row.names(Cstem3@IndicatorWeight)
temp3<-merge(Cstem3@IndicatorWeight,temp2,by=0,sort=FALSE)
Cstem3@IndicatorWeight[,4]<-temp3$Counts
c.3.set=sum(Cstem3@IndicatorWeight[,4])
Cstem3@IndicatorWeight[which(row.names(Cstem3@IndicatorWeight)=="000"),4]=35000-c.3.set
grid.newpage()
pdf(paste0(out.dir.name,"venn.pdf"))
plot(Cstem3)
dev.off()
}
#' GetResultsFromDiffBind
#'
#' Use DiffBind to process peak profiles
#'
#' @param mcf7
#' @param Mergereplicates
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#' output.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff_7_21_2017_2"
#' mcf7=resmcf
#' re<-GetResultsFromDiffBind(mcf7,"yes",output.file.dir)
#' re<-GetResultsFromDiffBind(mcf2,"yes",output.file.dir)
#'
GetResultsFromDiffBind<-function(mcf7,Mergereplicates=c("yes","no"),output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp<-mcf7
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
if(Mergereplicates=="yes"){
temp2<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp2)
##need to set the flexiable number identified from data sets
t <- temp2
temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
}else{
temp22<-temp
}
print(temp22)
Tissue1<-temp22$class[2,]
Tissue2<-unique(temp22$class[2,])
TF<-unique(temp22$class[3,])
TF.n<-length(TF)
temp3=file.path(output.file.dir,"Venn")
if(!dir.exists(temp3))
{
dir.create(temp3)
}
for(i in 1:length(Tissue2)){
po<-which(Tissue1 %in% Tissue2[i])
print(po)
if(length(po)==2)
{
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po,main=Tissue2[i])
dev.off()
}else if(length(po)==4){
po1<-po[c(1,3)]
po2<-po[c(2,4)]
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
dev.off()
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
dev.off()
}else
{
cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
}
}
p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
po<-which(Tissue1 %in% Tissue2[u])
if(length(po)==2)
{
common.peaks<-dba.overlap(temp22,mask=po)
y<-common.peaks$inAll
}else if(length(po)==4){
po1<-po[c(1,3)]
po2<-po[c(2,4)]
common.peaks.1<-dba.overlap(temp22,mask=po1)
y1<-common.peaks.1$inAll
common.peaks.2<-dba.overlap(temp22,mask=po2)
y2<-common.peaks.2$inAll
y<-list(y1=y1,y2=y2)
names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
}else{
y<-NULL}
y
},Tissue1,Tissue2,temp22)
names(p.common)<-Tissue2
p.common<-unlist(p.common,recursive = F)
p.common<-p.common[lapply(p.common,length) > 0]
if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
{
dir.create(file.path(output.file.dir,"common_peaks_bed"))
dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
}
#output common peaks to bed files
lapply(1:length(p.common),function(u,p.common,output.file.dir){
x=p.common[[u]]
x_name=names(p.common)[u]
df <- data.frame(seqnames=seqnames(x),
#starts=start(x)-1,
starts=start(x),
ends=end(x),
names=c(rep(".", length(x))),
scores=elementMetadata(x)[,1],
strands=strand(x))
#assign strand
df.str <- data.frame(seqnames=seqnames(x),
#starts=start(x)-1,
starts=start(x),
ends=end(x),
names=c(rep(".", length(x))),
scores=elementMetadata(x)[,1],
strands=c(rep(".", length(x))))
df.str.1<-df.str[-grep("random",df.str$seqnames),]
df.str.2<-df.str.1
df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
},p.common,output.file.dir)
AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
return(p.common)
}
#' GetSampleInfo
#'
#' GetSampleInfo
#'
#' @param input.sample.file
#' @param input.bam.file
#'
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/sorted_bam_files.txt'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny/sorted_bam_files_2.txt'
#'
#' re <- GetSampleInfo(input.sample.file,input.bam.file)
#'
#'
GetSampleInfo <- function(input.sample.file, input.bam.file)
{
re <- read.csv(input.sample.file)
re.c <- colnames(re)
bam.file <- read.table(input.bam.file, header = FALSE)
yy <- apply(bam.file, 1, function(u)
{
y <- basename(u)
pos <- regexpr("\\.", y)
pos <- pos - 1
y <- substr(y, 1, pos)
y
})
bam.file.sample.name <- cbind(bam.file, yy)
colnames(bam.file.sample.name) <- c("file.name", "ID")
Ab.type <- unique(as.character(re[, colnames(re) == re.c[3]]))
Cell.type <- unique(as.character(re[, colnames(re) == re.c[2]]))
re1 <- cbind(re[, 1:3], paste0(re[, 2], "_", re[, 3]))
re11 <- merge(re1, bam.file.sample.name, by = "ID")
colnames(re11)[4] = "Cell_TF"
chipseqCell <- function(n, es)
{
value <- list(name = n, es = es)
attr(value, "class") <- "ChipSeqCell"
value
}
cellType <- unique(as.character(re1[re1$Type_TF == "Input", ]$Type_Cell))
yy <- lapply(cellType, function(u, re11)
{
n <- nchar(u)
index1 <- which(substr(re11$Type_Cell, 1, n) == u)
z <- re11[index1, ]
index2 <- which(nchar(as.character(z$Type_Cell)) <= n + 2)
z2 <- z[index2, ]
cc <- chipseqCell(u, z2)
cc
}, re11)
colnames(re1)[4] = "Cell_TF"
Cell.Ab.type <- unique(as.character(re1[, 4]))
re2 <- lapply(1:length(Cell.Ab.type), function(u, Cell.Ab.type, re1)
{
x = Cell.Ab.type[u]
z = re1
ZZ <- as.character(z[which(z[, 4] == x), 1])
ZZ
}, Cell.Ab.type, re1)
names(re2) <- Cell.Ab.type
re21 <- re2[lapply(re2, length) == 1]
re22 <- re2[lapply(re2, length) == 2]
re3 <- list(re1 = re1, re2 = re2, re21 = re21, re22 = re22, re11 = re11,
y = yy)
return(re3)
}
#' peakcallwithinput
#'
#' @export
#' @example
#'
#' genome='Hs'
#' re <- peakcallwithinput(input.sample.file,input.bam.file,genome,output.dir,peakcaller)
#'
#'
peakcallwithinput <- function(input.sample.file, input.bam.file, genome = c("Hs",
"hs", "HS", "hS"), output.dir, peakcaller = c("macs14", "macs2"), peakPvalue)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
output.dir.name = dirname(input.sample.file)
temp3 = file.path(output.dir.name, output.dir)
if (!dir.exists(temp3))
{
dir.create(temp3)
}
peakcaller <- match.arg(peakcaller)
genome <- match.arg(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
PATH1 = Sys.getenv("PATH")
macs2_Lib = file.path("/nethome/axy148/NGS_tools/MACS/bin/")
Sys.setenv(PATH = paste0(macs2_Lib, ":", PATH1))
cmd1 <- Sys.which("macs2")[[1]]
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-p", peakPvalue, sep = " ")
})
cellInfo.run <- lapply(1:length(cellInfo), function(u, cellInfo, temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
x.run <- apply(x.sample, 1, function(x)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd12 = paste(cmd9, xx, "-c", x.input, cmd10, file.path(temp3, paste0(xx.name,
"_hs_1.00e-05_", peakcaller)), cmd11, sep = " ")
cmd12
})
x.run
}, cellInfo, temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
re <- list(cellInforun = cellInfo.run, zzz = zzz)
AnntationUsingChipSeeker(temp3, "peaks.bed", temp3, DD = 5000)
return(re)
}
select.sample <- c("MDA MB 231-DD-1_cJun", "MDA MB 231-1_cJun", "1833-1_cJun")
output.config.dir <- "~/"
configAndMultiplot <- function(res, select.sample, output.config.dir)
{
if (!dir.exists(output.config.dir))
{
dir.create(output.config.dir)
}
x <- res$re11
xx <- x[which(x$Cell_TF %in% select.sample), ]
xxx <- cbind.data.frame(xx$file.name, rep("-1", dim(xx)[1]), gsub(" ", "-",
xx$Cell_TF))
config.sample.name <- paste(gsub(" ", "-", select.sample), collapse = "-and-")
config.file.name <- file.path(output.config.dir, paste0(config.sample.name,
"-config.txt"))
write.table(xxx, file = config.file.name, col.names = FALSE, row.names = FALSE,
quote = FALSE, sep = "\t")
cmd0 <- "ngs.plot.r -G hg19 -R tss -C"
cmd1 <- "-O"
cmd2 <- "-L 4000 -RR 1 -CD 1 -CO \"blue\""
output.results <- file.path(output.config.dir, paste0(config.sample.name,
"results"))
cmd <- paste(cmd0, config.file.name, cmd1, output.results, cmd2, sep = " ")
system(cmd)
return(config.file.name)
}
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny/sorted_bam_files_2.txt'
#'
#' re <- ChipSeq:::matchBamInputGene(input.sample.file,input.bam.file,'$HOME/all_common_gene_unique.txt','$HOME/NgsConfigFile')
#'
#' R -e 'libraray(ChipSeq);re <- ChipSeq:::matchBamInputGene('/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv','/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt','$HOME/all_common_gene_unique.txt','$HOME/NgsConfigFile',,job.option = 'parallel')'
#'
#' bsub -P bbc -J 'bamPlot' -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::matchBamInputGene('/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv','/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt','~/all_common_gene_unique.txt','NgsConfigFile')'
#'
#' bsub -P bbc -J "bamPlot" -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::matchBamInputGene("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt","~/all_common_gene_unique.txt","NgsConfigFile3",add.input= "yes")'
matchBamInputGene <- function(input.sample.file, input.bam.file, input.gene.list,
output.dir, ngs.para = c("hg19", 4000, 1, 1, "total"), add.input = NULL)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u, add.input, cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
x.run <- apply(x.sample, 1, function(x, add.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
if (!is.null(add.input))
{
cmd12 = paste(paste0(xx, ":", x.input), input.gene.list, paste0(gsub(" ",
"-", Cell_TF), "_cJunAndp27"), sep = "\t")
} else
{
cmd12 = paste(xx, input.gene.list, paste0(gsub(" ",
"-", Cell_TF), "_cJunAndp27"), sep = "\t")
}
cat(cmd12, file = file.path(temp3, paste0(xx.name, "_config_cJunAndp27.txt")),
sep = "\t")
}, add.input, temp3)
x.run
}, add.input, cellInfo, temp3)
# dir.name=temp3 dir.name=reformatPath(dir.name)
file.name = file.path(temp3, dir(temp3, recursive = TRUE))
file.name.2 <- as.list(file.name)
# names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
zzz <- unlist(file.name.2)
lapply(1:length(zzz), function(u, zzz)
{
dir.name = dirname(zzz[u][[1]])
file_name = file_path_sans_ext(basename(zzz[u][[1]]))
cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
sep = " ")
# system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
cmd2 = cmd
cat(cmd2, "\n")
cat("\n")
system(cmd2)
}, zzz)
re <- list(cellInforun = cellInfo.run, zzz = zzz)
# AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
return(re)
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "bamCompare" -o %J.bamCompare.log -e %J.Compare.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCompare("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt","~/BamCompare")'
useBamCompare <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1",xx,"--bamfile2",x.input,sep=" ")
cmd2 <- "--binSize 25"
cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(x.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
sysyem("module unload python/2.7.3")
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
#' x <- ChipSeq:::usePegasus('parallel', Wall.time = '72:00',cores = 32,Memory = 16000,span.ptile = 16,'bamPlot')
usePegasus <- function(job.option = c("general", "parallel", "bigmem"), Wall.time,
cores, Memory, span.ptile, job.name, wait.job.name = NULL)
{
job.option <- match.arg(job.option)
switch(job.option, parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}, bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=", Memory,
"] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu", sep = " ")
}, general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=", Memory,
"] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu", sep = " ")
})
if (!is.null(wait.job.name))
{
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
} else
{
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.", job.name,
".log "), paste0("-e %J.", job.name, ".err -W"))
}
cmd = paste(cmd1, cmd0, sep = " ")
return(cmd)
}
#' IndexBamFile
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' input.file.pattern="*.bam"
#' index.file=9
#' output.file.dir="/scratch/projects/bbc/aiminy_project/
#' genome="Hs"
#'
#' IndexBamFile(input.file.dir,input.file.pattern,index.file,output.file.dir,genome)
#'
IndexBamFile <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
#library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
temp=Sys.time()
temp1=gsub(":","-",Sys.time())
temp2=gsub(" ","-",temp1)
temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
#print(file.name.2)
re.out<-file.name.2
cmd1="samtools index "
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd2=paste0(cmd1," ",x," ",paste0(temp3,"/",x_name,"_sorted"))
#print(cmd2)
system(cmd2)
},re.out,temp3)
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
#file.name.4 <-file.name.3[-1]
#re.out<-lapply(file.name.4,function(u){
# re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
# re
#})
#head(re.out[[1]])
# re.out.L<-lapply(re.out,function(u){
# re=length(u)
# #colnames(re)=c("Count","GeneName")
# re
# })
#
#
# # cmd="samtools sort"
# # input.file="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA/""2016-10-14-Hyunho-Yoon-16-1833-PF1502-1-p27_S4_R1.bam"
# # output.file="/scratch/projects/bbc/aiminy_project/Bam_marked_sorted/""2016-10-14-Hyunho-Yoon-16-1833-PF1502-1-p27_S4_R1.bam_sorted"
#
# if(genome=="Mm"){
#
# annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
#
# ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
#
# overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
#
# binOverFeature(overlaps, annotationData=annoData,
# radius=5000, nbins=20, FUN=length, errFun=0,
# ylab="count",
# main="Distribution of aggregated peak numbers around TSS")
#
# overlaps.trimmed<-trim(overlaps, use.names=TRUE)
#
# library(EnsDb.Mmusculus.v79)
# dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
# #seqinfo(dd.GRCm39.mm10)
# #seqlevels(dd.GRCm39.mm10)
#
# seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#
# seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
# #GRCm38/mm10
# #dd<-toGRanges(EnsDb.Mmusculus.v79)
# #seqinfo(dd)
# #library(ensembldb)
# #library(GenomeInfoDb)
# seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
# overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
# write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
# }else if(genome=="Hs"){
#
# library(EnsDb.Hsapiens.v75)
# #annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
#
# dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
#
# print(seqinfo(dd.hs))
# print(seqlevels(dd.hs))
#
# #print(seqlevels(dd.hs)[,1])
#
# #print(seqlevels(re.out[[1]])[,1])
#
# # seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# # "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# # "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#
# #temp4=
#
# re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
#
# x=re.out[[u]]
# x_name=names(re.out)[u]
#
# seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
# #print(seqinfo(re.out.trimmed))
# #print(seqlevels(re.out.trimmed))
# seqinfo(x)<-seqinfo(dd.hs)
# #GRCm38/mm10
# #dd<-toGRanges(EnsDb.Mmusculus.v79)
# #seqinfo(dd)
# #library(ensembldb)
# #library(GenomeInfoDb)
# seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
# re.out.trimmed<-trim(x, use.names=TRUE)
# overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
#
# write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
# },re.out,dd.hs)
#
# }
}
#' InstallRequiredPackage
#'
#' @return
#' @export
#'
#' @examples
#' InstallRequiredPackage()
#'
InstallRequiredPackage <- function() {
#install from bioc
source("http://bioconductor.org/biocLite.R")
biocLite("GenomicRanges")
biocLite("ChIPseeker")
biocLite("regioneR")
biocLite("DiffBind")
biocLite("BSgenome.Hsapiens.UCSC.hg19")
biocLite("TxDb.Mmusculus.UCSC.mm9.knownGene")
biocLite("EnsDb.Hsapiens.v75")
devtools::install_github("sheffien/simpleCache")
biocLite("LOLA")
biocLite("ChIPpeakAnno")
biocLite("BSgenome.Mmusculus.UCSC.mm9")
biocLite("motifStack")
biocLite("BSgenome.Ecoli.NCBI.20080805")
biocLite("EnsDb.Mmusculus.v75")
biocLite("EnsDb.Mmusculus.v79")
biocLite("BSgenome.Mmusculus.UCSC.mm10")
biocLite("org.Mm.eg.db")
#install from git
devtools::install_github("nsheff/LOLA")
#loading
library(BiocInstaller)
library(BiocInstaller)
library(ChIPseeker)
library(DiffBind)
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
library(TxDb.Mmusculus.UCSC.mm9.knownGene)
library(LOLA)
library("simpleCache")
library(ChIPpeakAnno)
library(rtracklayer)
library(org.Mm.eg.db)
library(motifStack)
library(BSgenome.Mmusculus.UCSC.mm10)
library(BSgenome.Mmusculus.UCSC.mm9)
library(EnsDb.Mmusculus.v75)
library(EnsDb.Mmusculus.v79)
library(ggplot2)
}
#' Title
#'
#' @return
#' @export
#'
#' @examples
#' LoadRequiredPackage()
#'
LoadRequiredPackage <- function() {
library(LOLA)
library(ChIPseeker)
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
library(clusterProfiler)
library(regioneR)
library(BSgenome.Hsapiens.UCSC.hg19)
}
#' ParserBamFile4NgsPlot
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#'
#' sh ~/Code/BashRunMACS1-4-2_4_Danny_chip_seq3.sh /scratch/projects/bbc/aiminy_project/Bam_sorted/ 7 ".sorted.bam$" /scratch/projects/bbc/aiminy_project/Bam_marked_sorted/
#'
#' dir.name="/scratch/projects/bbc/aiminy_project/Bam_sorted/"
#' index=7
#' input.file.pattern=".sorted.bam$"
#' out.dir.name="/scratch/projects/bbc/aiminy_project/Bam_marked_sorted/"
#' outfile="test.txt"
#'
#' re.from.bed.peng.4.venn<-ParserBamFile4NgsPlot(dir.name,index,input.file.pattern,out.dir.name,outfile)
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBamFile4NgsPlot<-function(dir.name,index,input.file.pattern,out.dir.name,outfile){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
#file.name="/scratch/projects/bbc/Project/Danny_chip/Alignment/BWA/2016-10-14-Hyunho-Yoon-02-MDA-MB-231-2-cJun_S5_R1.marked.bam"
#strsplit(file.name,split="\\/")
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index)
print(file.name.2)
ab<-names(file.name.2)
#ab<-"2016-10-14-Hyunho-Yoon-13-MDA-MB-231-DD-2-p27_S15_R1.marked.bam"
ab2<-sapply(strsplit(ab,split="\\."),"[[",1)
print(ab2)
#ab3<-sapply(strsplit(ab2,split="\\-"),"[[",length(strsplit(ab2,split="\\-")[[1]]))
re<-cbind(paste0(dir.name,names(file.name.2)),rep(-1,length(names(file.name.2))),ab2)
write.table(re, file =paste0(out.dir.name,outfile), append = FALSE, quote = F, sep = " ",eol = "\n", na = "NA", dec = ".", row.names = F, col.names = F)
cmd1="ngs.plot.r -G hg19 -R tss -C"
cmd2="-O"
cmd3="-L 4000"
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
cmd4=paste(cmd1,u,cmd2,u,cmd3,sep=" ")
print(cmd4)
system(cmd4, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
})
#sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
#names(re.out)=sample.name
# A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
# B=re.out[[2]] #10_WT_BM_ASXL1
# C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
# D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
#
# A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
# B.name=sample.name[2] #10_WT_BM_ASXL1
# C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
# D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#
# #A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
# #B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
# #class(A)
# #class(B)
#
#
# # TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
# # NumOverLap<-numOverlaps(A, B, count.once=TRUE)
# # pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
# # png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
# # plot(pt)
# # dev.off()
# #
# # lz <- localZScore(pt=pt, A=A, B=B)
# # png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
# # plot(lz)
# # dev.off()
# # }
# #
# # TestOverlap(A,B,A.name,B.name,out.dir.name,100)
# # TestOverlap(A,C,A.name,C.name,out.dir.name,100)
# # TestOverlap(A,D,A.name,D.name,out.dir.name,100)
# #
# # TestOverlap(B,C,B.name,C.name,out.dir.name,100)
# # TestOverlap(B,D,B.name,D.name,out.dir.name,100)
# # TestOverlap(C,D,C.name,D.name,out.dir.name,100)
# #
#
# # temp.name<-strsplit(names(file.name.2),split="\\.")
#
# #a = readBed(system.file("extdata", "examples/combined_regions.bed",
# # package="LOLA"))
#
#
# venn.plot <- venn.diagram(
# x = re.out[c(1,2)],
# filename = paste0(out.dir.name,names(re.out)[1],names(re.out)[2],"_overlap_venn.tiff"),
# height = 3000,
# width = 3500,
# resolution = 1000,
# col = "black",
# lty = "dotted",
# lwd = 1,
# fill = c("red","blue"),
# alpha = 0.50,
# label.col = c(rep("white",3)),
# cex = 0.5,
# fontfamily = "serif",
# fontface = "bold",
# cat.col = c("red","blue"),
# cat.cex = 0.5,
# cat.pos = 0.5,
# cat.dist = 0.05,
# cat.fontfamily = "serif"
# )
#
# venn.plot <- venn.diagram(
# x = re.out[c(1,3,5)],
# filename =paste0(out.dir.name,names(re.out)[1],names(re.out)[3],names(re.out)[5],"_overlap_venn.tiff"),
# col = "black",
# lty = "dotted",
# lwd = 2,
# fill = c("red", "orange", "blue"),
# alpha = 0.50,
# label.col = c(rep("white",7)),
# cex = 1,
# fontfamily = "serif",
# fontface = "bold",
# cat.col = c("red", "orange", "blue"),
# cat.cex = 0.8,
# cat.fontfamily = "serif"
# )
#return(re.out)
}
#' Title
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeakCall/"
#' input.file.pattern="*bam_mm_summits_from_PeakCall4Yang.bed"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#' output.test.file= "SMC1A_RAD21_overlap_test"
#'
#'
#' dir.name.2="/media/H_driver/2016/Yang/BedFromPeakCall_mm_100e_07/"
#' input.file.pattern.2="*_bam_mm_1_from_PeakCall4Yang.bed"
#'
#' out.dir.name.2="/media/H_driver/2016/Yang/Results/"
#' output.test.file.2= "SMC1A_RAD21_mm_100e-07_peaks_overlap_test"
#'
#' ParserBedFile(dir.name,input.file.pattern,out.dir.name)
#'
#' re.mm.100e.07<-ParserBedFile(dir.name.2,input.file.pattern.2,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_2.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_2.Rhistory"))
#'
ParserBedFile<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.3,function(u){
re=readBed(u)
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(sapply(strsplit(names(re.out),split="_i"),"[[",1),split="2470IUPUI_"),"[[",2)
A=re.out[[2]] #SMC1
B=re.out[[4]] #Rad21
C=re.out[[1]] #ASXL1
A.name=sample.name[2] #SMC1 name
B.name=sample.name[4] #Rad21 name
C.name=sample.name[1] #ASXL1 name
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
TestOverlap <- function(A, B, A.name,B.name,out.dir.name) {
NumOverLap<-numOverlaps(A, B, count.once=TRUE)
pt <- overlapPermTest(A=A, B=B, ntimes=50)
png(filename=paste0(out.dir.name,A.name,B.name,"_overlap_test.png"))
plot(pt)
dev.off()
lz <- localZScore(pt=pt, A=A, B=B)
png(filename=paste0(out.dir.name,A.name,B.name,"_localZScore.png"))
plot(lz)
dev.off()
}
TestOverlap(A,B,A.name,B.name,out.dir.name)
TestOverlap(A,C,A.name,C.name,out.dir.name)
TestOverlap(B,C,B.name,C.name,out.dir.name)
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
return(re.out)
}
#' ParserBedFile4Peng
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeng/"
#' input.file.pattern="*.bed"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng<-ParserBedFile4Peng(dir.name,input.file.pattern,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4Peng<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
re=readBed(u)
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
B=re.out[[2]] #10_WT_BM_ASXL1
C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
B.name=sample.name[2] #10_WT_BM_ASXL1
C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
NumOverLap<-numOverlaps(A, B, count.once=TRUE)
pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
plot(pt)
dev.off()
lz <- localZScore(pt=pt, A=A, B=B)
png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
plot(lz)
dev.off()
}
TestOverlap(A,B,A.name,B.name,out.dir.name,100)
TestOverlap(A,C,A.name,C.name,out.dir.name,100)
TestOverlap(A,D,A.name,D.name,out.dir.name,100)
TestOverlap(B,C,B.name,C.name,out.dir.name,100)
TestOverlap(B,D,B.name,D.name,out.dir.name,100)
TestOverlap(C,D,C.name,D.name,out.dir.name,100)
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
return(re.out)
}
#' ParserBedFile4PengVenn
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeng/"
#' input.file.pattern="*combine_chr_st_end.bed"
#'
#' input.file.pattern="*combine_chr_st.bed"
#'
#'
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng.4.venn<-ParserBedFile4PengVenn(dir.name,input.file.pattern,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4PengVenn<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
re=read.table(u,header=FALSE)
re<-as.character(re[,1])
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
names(re.out)=sample.name
A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
B=re.out[[2]] #10_WT_BM_ASXL1
C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
B.name=sample.name[2] #10_WT_BM_ASXL1
C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
# TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
# NumOverLap<-numOverlaps(A, B, count.once=TRUE)
# pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
# png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
# plot(pt)
# dev.off()
#
# lz <- localZScore(pt=pt, A=A, B=B)
# png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
# plot(lz)
# dev.off()
# }
#
# TestOverlap(A,B,A.name,B.name,out.dir.name,100)
# TestOverlap(A,C,A.name,C.name,out.dir.name,100)
# TestOverlap(A,D,A.name,D.name,out.dir.name,100)
#
# TestOverlap(B,C,B.name,C.name,out.dir.name,100)
# TestOverlap(B,D,B.name,D.name,out.dir.name,100)
# TestOverlap(C,D,C.name,D.name,out.dir.name,100)
#
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
venn.plot <- venn.diagram(
x = re.out[c(1,2)],
filename = paste0(out.dir.name,names(re.out)[1],names(re.out)[2],"_overlap_venn.tiff"),
height = 3000,
width = 3500,
resolution = 1000,
col = "black",
lty = "dotted",
lwd = 1,
fill = c("red","blue"),
alpha = 0.50,
label.col = c(rep("white",3)),
cex = 0.5,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","blue"),
cat.cex = 0.5,
cat.pos = 0.5,
cat.dist = 0.05,
cat.fontfamily = "serif"
)
venn.plot <- venn.diagram(
x = re.out[c(1,3,5)],
filename =paste0(out.dir.name,names(re.out)[1],names(re.out)[3],names(re.out)[5],"_overlap_venn.tiff"),
col = "black",
lty = "dotted",
lwd = 2,
fill = c("red", "orange", "blue"),
alpha = 0.50,
label.col = c(rep("white",7)),
cex = 1,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red", "orange", "blue"),
cat.cex = 0.8,
cat.fontfamily = "serif"
)
return(re.out)
}
#' ParserBedFile4PengXls
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/MACS/MACS/"
#' input.file.pattern="*.xls"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng.Xls<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' re.from.bed.peng.Xls.2<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' re.from.bed.peng.Xls.3<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4PengXls<-function(dir.name,input.file.pattern,out.dir.name){
dir.name=reformatPath(dir.name)
out.dir.name=reformatPath(out.dir.name)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",8)
print(file.name.2)
re.out<-lapply(names(file.name.2),function(u){
#print(names(u))
uu=file.name.2[[u]]
re=read.table(uu,header=TRUE)
colnames(re)[c(7,9)]=c("-10*LOG10(pvalue)","FDR(%)")
temp<-re
temp$start<-temp$start-1
temp$end<-temp$end-1
temp$summit<-temp$summit-1
temp
summitPeak<-temp$start+temp$summit
temp2<-temp
temp2$start<-summitPeak-49
temp2$end<-summitPeak+50
temp2$summit<-summitPeak
temp2$length<-temp2$end-temp2$start+1
temp2
rownames(temp2)<-paste0("MACS_peak_",rownames(temp2))
temp3<-toGRanges(temp2)
names(temp3)<-rownames(temp2)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Mmusculus.UCSC.mm10")
genome(temp3.trimmed)<-"mm10"
seq.temp3<-getAllPeakSequence(temp3.trimmed,genome=genome.mm10)
write2FASTA(seq.temp3, paste0(out.dir.name,u,".fa"))
re2<-list(originalPeak=re,down1Peak=temp,aroundSummit100Peak=temp2,GR=temp3)
re2
})
sample.name<-sapply(strsplit(names(file.name.2),split="_peaks_"),"[[",1)
names(re.out)<-sample.name
return(re.out)
}
#' ParserReadFiles
#'
#' @param input.file.dir
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="~/TestBam/"
#' out.file.dir="~/"
#' testR<-ParserReadFiles(input.file.dir)
#'
#'
ParserReadFiles <- function(input.file.dir,input.file.type) {
#For input
dir.name=input.file.dir
dir.name=reformatPath(dir.name)
file.name=file.path(dir.name,dir(dir.name,recursive = TRUE))
file.name.2<-as.list(file.name)
file.name.3<-lapply(1:length(file.name.2),function(u,input.file.type,file.name.2){
tmp=file.name.2
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
n <- length(grep(input.file.type,file_name))
#if(file_ext(file_name)==input.file.type)
if(n == 1) {
re<-file.path(path_name,file_name)
#names(re)[u]<-file_name
}else
{re<-NULL}
re
},input.file.type,file.name.2)
file.name.4<-file.name.3[lapply(file.name.3, length) > 0]
names(file.name.4)=unlist(lapply(1:length(file.name.4),function(u,file.name.4){
tmp=file.name.4
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
file_name
},file.name.4))
#print(file.name.4)
#For output
#output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",temp)
#temp2=gsub(" ","-",temp1)
#temp3=file.path(output.dir.name,"ReadBam_at_",temp2)
#temp3=output.dir.name
#dir.create(temp3)
re2<-list(input=file.name.4)
return(re2)
}
#' ChipSeq:::getGeneBedName("/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny","peaks.csv","/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny")
#'
getGeneBedName <- function(input.file.dir,file.pattern,output.file.dir) {
re<-ParserReadFiles(input.file.dir,file.pattern)
file.name.2<-re$input
re.out<-file.name.2
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp3 =output.file.dir
cmd.1 <- lapply(1:length(re.out),function(u,re.out,temp3){
x <- read.csv(re.out[[u]],header = TRUE)
xx <- x[grep("Promoter",x$annotation),]
chr <- which(colnames(xx) == "seqnames")
gs <- which(colnames(xx) == "geneStart")
ge <- which(colnames(xx) == "geneEnd")
file_name = file_path_sans_ext(basename(re.out[[u]]))
write.table(xx[,c(chr,gs,ge)],file=file.path(temp3,paste0(file_name,"_gene.bed")),quote = F,col.names = F,row.names = F,sep="\t")
write.table(unique(xx$SYMBOL),file=file.path(temp3,paste0(file_name,"_gene_name.txt")),quote = F,col.names = F,row.names = F)
ps <- which(colnames(xx) == "start")
pe <- which(colnames(xx) == "end")
write.table(xx[,c(chr,ps,pe)],file=file.path(temp3,paste0(file_name,"_peaks.bed")),quote = F,col.names = F,row.names = F,sep="\t")
},re.out,temp3)
}
#' PeakCallAndAnnotation
#'
#' Call peak for bam files using macs14 and perform peak annotation using ChIPpeakAnno
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' input.file.pattern="*.bam"
#' index.file=9
#' output.file.dir="/scratch/projects/bbc/aiminy_project/
#' genome="Hs"
#'
#' PeakCallAndAnnotation(input.file.dir,output.file.dir,genome)
#'
PeakCallAndAnnotation <- function(input.file.dir,output.file.dir,genome) {
re<-ParserReadFiles(input.file.dir,"bam",output.file.dir)
file.name.2<-re$input
output.dir.name=re$output
temp3=paste0(output.dir.name,"_PeakCall")
dir.create(temp3)
re.out<-file.name.2
cmd9="macs14 -t "
cmd10="-f BAM -g hs -n "
cmd11=" -m 6,18 --bw=200 -p 0.00001"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd12=paste(cmd9,x,cmd10,paste0(temp3,"/",paste0(x_name,"_hs_1.00e-05_macs142")),cmd11,sep=" ")
print(cmd12)
system(cmd12, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
#AnnotatePeak2(paste0(temp3,"/"),"*macs142_peaks.bed",7,paste0(output.dir.name,"PeakAnnotation_at_",temp2),genome="Hs")
AnnotatePeak3(paste0(temp3,"/"),paste0(output.dir.name,"_PeakAnnotation"),
genome="Hs")
BamFileSortIndexVisualization2(re,genome)
}
BamFileSortIndexVisualization2 <- function(input,genome) {
#library(ChIPpeakAnno)
re<-input
file.name.2<-re$input
output.dir.name=re$output
temp3=paste0(output.dir.name,"_visualization")
dir.create(temp3)
re.out<-file.name.2
cmd1="samtools sort"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd2=paste0(cmd1," ",x," ",paste0(temp3,"/",x_name,"_sorted"))
#print(cmd2)
system(cmd2)
},re.out,temp3)
cmd3="samtools index"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd4=paste0(cmd3," ",paste0(temp3,"/",x_name,"_sorted.bam"))
#print(cmd2)
system(cmd4)
},re.out,temp3)
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd8=paste(cmd5,paste0(temp3,"/",paste0(x_name,"_sorted.bam")),cmd6,paste0(temp3,"/",paste0(x_name,"_sorted")),cmd7,sep=" ")
print(cmd8)
system(cmd8, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
}
#' ProcessUsingCHIPpeakAnno
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#' input.file.dir="/media/H_driver/2016/Yang/MACS/MACS/"
#' input.file.pattern="*.bed"
#' output.file.dir="/media/H_driver/2016/Yang/MACS/MACS/"
#'
#' ProcessUsingCHIPpeakAnno(input.file.dir,input.file.pattern,output.file.dir)
#'
ProcessUsingCHIPpeakAnno <- function(input.file.dir,input.file.pattern,output.file.dir) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
dir.create(output.dir.name)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",8)
print(file.name.2)
file.name.3<-file.name.2
sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
names(file.name.3)=sample.name
file.name.4 <-file.name.3[-1]
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
annoData[1:2]
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
re<-makeVennDiagram(re.out[c(2,4,1)],NameOfPeaks=c("SMC1A", "RAD21","ASXL1"),totalTest=35000)
#fisher exact test
UseFisher <- function(temp.ct,index.A,index.B,totalN) {
total.peaks=totalN
A=sum(temp.ct[which(temp.ct[,index.A]==1&temp.ct[,index.B]==1),4])
B=sum(temp.ct[which(temp.ct[,index.A]==1&temp.ct[,index.B]==0),4])
C=sum(temp.ct[which(temp.ct[,index.A]==0&temp.ct[,index.B]==1),4])
D=total.peaks-(A+B+C)
ctb<-matrix(c(A,B,C,D),nrow = 2,dimnames =list(c("In", "Not"),c("In", "Not")))
#re<-fisher.test(ctb)
print(ctb)
re.fisher<-fisher.test(ctb, alternative='greater')[c("p.value","estimate")]
re.fisher
}
temp.ct<-ol$venn_cnt
#A vs B
index.A<-grep("SMC1",colnames(temp.ct))
index.B<-grep("Rad21",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.AB=tempRe$p.value
OR.fisher.AB=tempRe$estimate
#A vs C
index.A<-grep("SMC1",colnames(temp.ct))
index.B<-grep("ASXL1",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.AC=tempRe$p.value
OR.fisher.AC=tempRe$estimate
#B vs C
index.A<-grep("Rad21",colnames(temp.ct))
index.B<-grep("ASXL1",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.BC=tempRe$p.value
OR.fisher.BC=tempRe$estimate
pVal.fisher.AB
OR.fisher.AB
pVal.fisher.AC
OR.fisher.AC
pVal.fisher.BC
OR.fisher.BC
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
library(TxDb.Hsapiens.UCSC.mm9.knownGene)
aCR<-assignChromosomeRegion(overlaps, nucleotideLevel=FALSE,
precedence=c("Promoters", "immediateDownstream",
"fiveUTRs", "threeUTRs",
"Exons", "Introns"),
TxDb=TxDb.Mmusculus.UCSC.mm9.knownGene)
barplot(aCR$percentage, las=3)
pie1(aCR$percentage,las=3)
dc<-annoGR(TxDb.Mmusculus.UCSC.mm9.knownGene)
seqinfo(dc)
seqlevels(dc)
seqinfo(overlaps)
seqlevels(overlaps)
#GRCm38/mm10
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
seqinfo(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
seqinfo(overlaps)
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
#GRCm38/mm10
dd<-toGRanges(EnsDb.Mmusculus.v79)
seqinfo(dd)
library(ensembldb)
library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
library(org.Mm.eg.db)
overlaps.anno.with.entrez.id <- addGeneIDs(overlaps.anno,"org.Mm.eg.db",IDs2Add = "entrez_id")
write.csv(as.data.frame(unname(overlaps.anno.with.entrez.id)), paste0(out.dir.name,"other_anno.csv"))
pie1(table(overlaps.anno.with.entrez.id$insideFeature))
library("DBI")
over <- getEnrichedGO(overlaps.anno.with.entrez.id, orgAnn="org.Mm.eg.db",
maxP=.05, minGOterm=10,
multiAdjMethod="BH", condense=TRUE)
# over.gene.symbol <- getEnrichedGO(overlaps.anno.with.entrez.id, orgAnn="org.Mm.eg.db",
# feature_id_type="gene_symbol",
# maxP=.05, minGOterm=10,
# multiAdjMethod="BH",condense=TRUE)
head(over[["bp"]][, -c(3, 10)])
library(org.Hs.eg.db)
e2s = toTable(org.Mm.egSYMBOL)
tempDS=over$bp
tempDS2<-data.frame(apply(tempDS,1,function(u,e2c){
#print(u[1])
x=u[11]
tempId<-unlist(strsplit(as.character(x),split=";"))
index<-which(!is.na(match(e2s[,1],tempId)))
index<-match(tempId,e2s[,1])
geneS<-paste(e2s[index,2], collapse=";")
geneS
#print(geneS)
},e2c))
tempDS3<-cbind(tempDS,tempDS2)
colnames(tempDS3)[12]="GeneSymbol"
Draw4GO <- function(tempDS3) {
x=tempDS3
y=x[order(x$pvalue,decreasing = TRUE),]
z=y[1:10,c(1,3,4,5,6,10)]
Function<-z$Definition
negative_log10p=-log10(z$pvalue)
library(ggplot2)
#ggplot(z, aes(x=z$go.id, y=negative_log10p,fill=factor(z$go.id)))+geom_bar(stat="identity")+geom_hline(yintercept = -log10(0.05))+coord_flip()
ggplot(z, aes(go.id,pvalue, fill = go.id)) +
geom_bar(stat="identity")+ scale_x_discrete(labels=z$count.InDataset, limits=factor(z$go.id))+ scale_fill_discrete(breaks = z$go.id,
name="GO term")+theme(legend.text = element_text(colour="black", size = 11, face = "bold"))+
theme(axis.text.x = element_text(angle = 90, hjust = 1))+ggtitle("GO enrichment analysis")+labs(x="Gene Counts",y="-log p-value")
}
write.table(tempDS3,file=paste0(out.dir.name,"BP_.txt"),row.names = FALSE,quote=FALSE,sep="\t")
#anno <- annoGR(EnsDb.Hsapiens.v79)
ree<-annotatePeakInBatch(overlaps,
AnnotationData=dc,
output="nearestBiDirectionalPromoters",
bindingRegion=c(-2000, 500))
ree2 <- addGeneIDs(ree,
"org.Mm.eg.db",
IDs2Add = "entrez_id")
re<-makeVennDiagram(re.out[c(2,4,1)],NameOfPeaks=c("SMC1A", "RAD21","ASXL1"),totalTest=35000)
library(BSgenome.Mmusculus.UCSC.mm9)
upseqs<-Views(Mmusculus,overlaps)
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
mm9.S<-Seqinfo(genome="mm9")
seqinfo(overlaps,force=TRUE) <- Seqinfo(genome="mm9")
seqlevels(mm9.S) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps) <- mm9.S
goodGR <- trim(overlaps)
overlaps.trimmed<-goodGR
seq<-getAllPeakSequence(overlaps.trimmed,genome=Mmusculus)
seqs.mm9 <- getSeq(Mmusculus,overlaps.trimmed)
write2FASTA(seq, paste0(out.dir.name,"WT_triple.fa"))
## We can also try simulation data
seq.sim.motif <- list(c("t", "g", "c", "a", "t", "g"),
c("g", "c", "a", "t", "g", "c"))
set.seed(1)
seq.sim <- sapply(sample(c(2, 1, 0), 1000, replace=TRUE, prob=c(0.07, 0.1, 0.83)),
function(x){
s <- sample(c("a", "c", "g", "t"),
sample(100:1000, 1), replace=TRUE)
if(x>0){
si <- sample.int(length(s), 1)
if(si>length(s)-6) si <- length(s)-6
s[si:(si+5)] <- seq.sim.motif[[x]]
}
paste(s, collapse="")
})
os <- oligoSummary(seq, oligoLength=6, MarkovOrder=3,
quickMotif=TRUE)
zscore <- sort(os$zscore, decreasing=TRUE)
h <- hist(zscore, breaks=100, main="Histogram of Z-score")
text(zscore[1:2], rep(5, 2),
labels=names(zscore[1:2]), adj=0, srt=90)
pfms <- mapply(function(.ele, id)
new("pfm", mat=.ele, name=paste("SAMPLE motif", id)),
os$motifs, 1:length(os$motifs))
motifStack(pfms[[1]])
motifStack(pfms[[2]])
motifStack(pfms[[3]])
motifStack(pfms[[4]])
}
# input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
# input.file.pattern="*macs142_peaks.bed"
# output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#
# AnnotatePeak(input.file.dir,input.file.pattern,8,output.file.dir,genome="Mm")
# AnnotatePeak(input.file.dir,input.file.pattern,7,output.file.dir,genome="Hs")
AnnotatePeak <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
temp=Sys.time()
temp1=gsub(":","-",Sys.time())
temp2=gsub(" ","-",temp1)
temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3[-1]
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
ProcessUsingLOLA <- function() {
#dbPath = system.file("extdata", "hg19", package="LOLA")
dbPath = "/media/H_driver/2016/Yang/Results/data/groups/lab_bock/shared/resources/regions/LOLACore/mm10"
regionDB = loadRegionDB(dbLocation=dbPath)
#read triple overlap peaks
regionSetA = readBed("/media/H_driver/2016/Yang/Results/WT_triple_overlap.bed")
#read each peak
regionSet.WT.ASXL1=readBed("/media/H_driver/2016/Yang/BedFromPeng/10_WT_BM_ASXL1_peaks_from_PeakCall4Yang.bed")
regionSet.WT.SMC1=readBed("/media/H_driver/2016/Yang/BedFromPeng/11_2470IUPUI_WT_BM_SMC1_peaks_from_PeakCall4Yang.bed")
regionSet.WT.Rad21=readBed("/media/H_driver/2016/Yang/BedFromPeng/13_2470IUPUI_WT_BM_Rad21_peaks_from_PeakCall4Yang.bed")
db.841<-regionDB$regionGRL
userSets = GRangesList(regionSet.WT.ASXL1,regionSet.WT.SMC1,regionSet.WT.Rad21)
db.841.2<-c(db.841,userSets)
userUniverse<-buildRestrictedUniverse(userSets)
userUniverse.844<-buildRestrictedUniverse(db.841.2)
re.SMC1A.RAD21.only<-makeVennDiagram(re.out[c(2,4)],NameOfPeaks=c("SMC1A", "RAD21"),totalTest=35000)
ol.SMC1A.RAD21.only <- findOverlapsOfPeaks(re.out[c(2,4)])
ol.SMC1A.RAD21.only.2<-ol.SMC1A.RAD21.only$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed`
checkUniverseAppropriateness(ol.SMC1A.RAD21.only.2,userUniverse.844, cores = 1,fast = FALSE)
locResults2 = runLOLA(userSets,userUniverse,regionDB, cores=1)
locResults = runLOLA(regionSetA,userUniverse,regionDB, cores=1)
#SMC1A and RAD21 overlap
locResults.SMC1A.RAD21 = runLOLA(ol.SMC1A.RAD21.only.2,userUniverse,regionDB, cores=4)
head(locResults.SMC1A.RAD21)
pValue<-as.data.frame(10^(-locResults.SMC1A.RAD21$pValueLog))
temp<-as.data.frame(locResults.SMC1A.RAD21)
locResults.SMC1A.RAD21.with.p<-cbind(temp[,1:3],pValue,temp[,4:23])
colnames(locResults.SMC1A.RAD21.with.p)[4]="pValue"
#It takes extensive computation resource, be carefully to run
locResults.SMC1A.RAD21.based.large.universe = runLOLA(ol.SMC1A.RAD21.only.2,userUniverse.844,regionDB, cores=4)
write.table(locResults.SMC1A.RAD21.with.p,
file=paste0(out.dir.name,"SMC1A.RAD21.with.Other.Antibody.csv"),sep=",",
quote = FALSE,row.names = FALSE,col.names = TRUE)
write.table(locResults,file=paste0(out.dir.name,"Other.csv"),sep=",",quote = FALSE,row.names = FALSE,col.names = TRUE)
listRegionSets(regionDB)
extractEnrichmentOverlaps(locResults,regionSetA, regionDB)
loadCaches("/media/H_driver/2016/Yang/Results/LOLACoreCaches_latest.tgz")
save.image(file =paste0(out.dir.name,"re_search.RData"))
# regionSetB = readBed("lola_vignette_data/setB_100.bed")
# regionSetC = readBed("lola_vignette_data/setC_100.bed")
# activeDHS = readBed("lola_vignette_data/activeDHS_universe.bed")
# data("sample_universe", package="LOLA")
#
# dbPath = system.file("extdata", "hg19", package="LOLA")
# regionDB = loadRegionDB(dbLocation=dbPath)
# data("sample_universe", package="LOLA")
# data("sample_input", package="LOLA")
#
# getRegionSet(regionDB, collections="ucsc_example", filenames="vistaEnhancers.bed")
# getRegionSet(dbPath, collections="ucsc_example", filenames="vistaEnhancers.bed")
#
# res = runLOLA(userSets, userUniverse, regionDB, cores=1)
# locResult = res[2,]
}
#' parser4diffbind
#'
#' parser4diffbind
#'
#' @param input.sample.file
#' @param input.bed.dir
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.sample.file = "/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv"
#' input.bed.dir = "/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/"
#' input.file.pattern = "bed"
#' output.dir="BindDiff_4_7_2017-cutoff_10k_test"
#' output.dir="/Volumes/Bioinformatics$/2017/DannyNewData/testRun"
#' resmcf<-parser4diffbind(input.sample.file,input.bed.dir,input.file.pattern,output.dir)
#'
parser4diffbind<-function(input.sample.file,input.bed.dir,input.file.pattern,output.dir,
summit_peak=NULL){
sample.info<-read.csv(input.sample.file,header = TRUE)
sample.info.2 <- sample.info[,-dim(sample.info)[2]]
out.dir.name = dirname(input.sample.file)
re<-ParserReadFiles(input.bed.dir,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2<-FL(re.bed,'peaks')
if(!is.null(summit_peak)){
re.summits.only.bed<-FL(re.bed,'summits')
}
#put peak files into database
re.peaks.only.bed.3<-list_to_df(re.peaks.only.bed.2)
print(re.peaks.only.bed.3)
pos <- regexpr('R1', re.peaks.only.bed.3$ID)
ID2 <- substr(re.peaks.only.bed.3$ID,1,pos+1)
re.peaks.only.bed.3 <- cbind(ID2,re.peaks.only.bed.3)
colnames(re.peaks.only.bed.3)=c("ID","ID2")
re.peaks.only.bed.4<-merge(sample.info.2,re.peaks.only.bed.3,by="ID")
replicate<-apply(re.peaks.only.bed.4,1,function(x){
ID=as.character(x[1])
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
p1=nchar(ID)+2
p2=regexpr(TF,ID2)
p2=p2-2
xx=substr(ID2,p1,p2)
cell=substr(xx,1,nchar(xx)-2)
rep=substr(xx,nchar(xx),nchar(xx))
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
z
})
res <- do.call(rbind.data.frame,replicate)
colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
re.peaks.only.bed.5 <- res
mcf<-NULL
for(i in 1:dim(re.peaks.only.bed.5)[1]){
#peaks.v=readBed(as.character(re.peaks.only.bed.5[i,6]))
peaks.v <- readPeakFile(as.character(re.peaks.only.bed.5[i,6]), as = "GRanges")
peak.caller.v="macs142"
sampID.v=as.character(re.peaks.only.bed.5[i,1])
sampID.v=substr(sampID.v,1,nchar(sampID.v)-5)
tissue.v=as.character(re.peaks.only.bed.5[i,2])
factor.v=as.character(re.peaks.only.bed.5[i,3])
condition.v=as.character(re.peaks.only.bed.5[i,4])
replicate.v=as.numeric(as.character(re.peaks.only.bed.5[i,5]))
if(is.null(mcf)){
mcf <- dba.peakset(NULL,peaks=peaks.v,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}else{
mcf <- dba.peakset(mcf,peaks=peaks.v,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}
}
print(mcf)
temp3=output.dir
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
save(mcf,file=file.path(temp3,"mcf.RData"))
pdf(file.path(temp3,"corrmap.pdf"))
dba.plotHeatmap(mcf,margin=23)
dev.off()
GetResultsFromDiffBind(mcf,"yes",temp3)
return(mcf)
}
#' Title
#'
#' @param dir.name
#'
#' @return
#' @export
#'
#' @examples
#' dir.name="/media/H_driver/2016/Yang/MACS/MACS/"
#' reformatPath(dir.name)
reformatPath <- function(dir.name){
CheckOPS=Sys.info()[['sysname']]
if(CheckOPS=="Darwin"){
if(length(grep('H_driver',dir.name))!=0){
temp=unlist(strsplit(dir.name,split="\\/"))
temp[2]="Volumes"
temp[3]="Bioinformatics$"
dir.name=paste0(paste0(temp,collapse = "/"),"/")
}
}
return(dir.name)
}
#' useHOMER
#'
#' @return
#' @export
#'
#' @examples
#' res <- useHOMER("findMotifs.pl")
#' system(res)
#'
useHOMER <- function(function.name){
R_LIB=.libPaths()
PATH1=Sys.getenv("PATH")
homer_Lib=file.path(R_LIB,"ChipSeq/homer/bin")
Sys.setenv(PATH=paste0(homer_Lib,":",PATH1))
cmd1 <- Sys.which(function.name)[[1]]
PATH2=Sys.getenv("PATH")
cat(PATH1,"\n")
cat(PATH2,"\n")
return(cmd1)
}
# Filtering list based on certain pattern of the names of list elements
FL <- function(re.bed, patt) {
re.peaks.only.bed<-lapply(1:length(re.bed),function(u,re.bed,patt){
tmp=re.bed
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
#pos_summits=regexpr('summits',file_name)
pos_peaks=regexpr(patt,file_name)
if(pos_peaks>0){
y<-x
}else{
y<-NULL
}
y
},re.bed,patt)
re.peaks.only.bed.2<-re.peaks.only.bed[lapply(re.peaks.only.bed,length) > 0]
names(re.peaks.only.bed.2)=unlist(lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2){
tmp=re.peaks.only.bed.2
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
file_name
},re.peaks.only.bed.2))
return(re.peaks.only.bed.2)
}
list_to_df <- function(list_for_df) {
list_for_df <- as.list(list_for_df)
nm <- names(list_for_df)
if (is.null(nm))
nm <- seq_along(list_for_df)
ID<-sapply(1:length(nm),function(u,nm){
x=nm[u]
pos=regexpr("\\.",x)
pos1=pos-1
y<-substr(x,1,pos1)
y
},nm)
df <- data.frame(ID=ID,name = nm, stringsAsFactors = FALSE)
df$value <- unname(list_for_df)
df
}
get_os <- function(){
sysinf <- Sys.info()
if (!is.null(sysinf)){
os <- sysinf['sysname']
if (os == 'Darwin')
os <- "osx"
} else { ## mystery machine
os <- .Platform$OS.type
if (grepl("^darwin", R.version$os))
os <- "osx"
if (grepl("linux-gnu", R.version$os))
os <- "linux"
}
tolower(os)
}
#' plotBam2
#'
#' @export
#' @example
#'
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny/Bam_files.txt'
#'
#' output.dir <- '/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny/BamMerged'
#'
#' re <- plotBam2(input.sample.file,input.bam.file,output.dir)
#'
#'
plotBam2 <- function(input.sample.file,input.bam.file,output.dir)
{
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$re11
cell.name <- unique(unlist(lapply(cellInfo$Type_Cell,function(u){substr(as.character(u),1,nchar(as.character(u))-2)})))
cmd="samtools merge"
cmd.3 <- lapply(cell.name,function(u,cellInfo,output.dir){
cat(u,"\n")
x <- cellInfo[which(substr(as.character(cellInfo$Type_Cell),1,nchar(as.character(cellInfo$Type_Cell))-2)==u),]
x$Cell_TF <- gsub(" ","-",x$Cell_TF)
tf <- as.character(x$Type_TF)
tf.unique <- unique(tf)
y <- combn(1:length(tf),2,function(x)list(c(x[1],x[2])))
yy <- lapply(tf.unique,function(u,tf,y){
x <- which(tf %in% u)
x
},tf,y)
yyy <- setdiff(y,yy)
yyyy <- yyy[lapply(yyy, length) > 0]
cmd.2 <- lapply(yyyy,function(u,x,output.dir){
cmd.1 <- paste(cmd,file.path(output.dir,paste0(paste(x[c(u),]$Cell_TF,collapse ="-"),"_merged.bam")),paste(x[c(u),]$file.name,collapse = " "),collapse = " ")
},x,output.dir)
cmd.2
},cellInfo,output.dir)
cmd.4 <- unlist(cmd.3)
mergeBamFilesUseJobArray(cmd.4)
}
mergeBamFilesUseJobArray <- function(cmd)
{
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
total <- system('echo $LSB_JOBINDEX_END',intern = TRUE)
cat(index,"\n\n")
cat(total,"\n\n")
u <- as.integer(index)
system(cmd[u],intern = TRUE)
cat(cmd,"\n\n")
}
#' R -e 'library(DoGs);library(ChipSeq);ChipSeq:::runPlotBam2("~/Danny/SampleID_INFO_ChIP_new_Danny.csv","~/Danny/Alignment/BWA/bam_files.txt","/scratch/projects/bbc/aiminy_project/DannyBamMerged")'
runPlotBam2 <-
function(input.sample.file,
input.bam.file,
output.dir) {
# This setting works
Rfun1 <-
'library(DoGs);library(ChipSeq);re <- ChipSeq:::plotBam2('
input1 <- input.sample.file
input2 <- input.bam.file
output <- output.dir
Rinput <-
paste0('\\"',
input1,
'\\",',
'\\"',
input2,
'\\",',
'\\"',
output,
'\\"')
Rfun2 <- ')'
Rfun3 <- paste0(Rfun1, Rinput, Rfun2)
n.job <- 12
mergeBam <-
DoGs:::createBsubJobArrayRfun(Rfun3, paste0("mergeBam[1-", n.job, "]"), NULL)
system(mergeBam)
}
#' bsub -P bbc -J "bamPlot" -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::plotMergedBamWgene("/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot4Merged","*_sorted.bam","~/all_common_gene_unique.txt","NgsPlot4Merged")'
#'
plotMergedBamWgene <-
function(input.bam.file.dir,
file.pattern,
input.gene.list,
output.dir,
ngs.para = c("hg19", 4000, 1, 1, "total"),
add.input = NULL)
{
cellInfo <-
list.files(
input.bam.file.dir,
pattern = file.pattern,
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE
)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
cellInfo.run <- lapply(1:length(cellInfo), function(u, cellInfo,
temp3)
{
xx <- cellInfo[u]
xxx <- file_path_sans_ext(basename(xx))
cmd12 = paste(xx, input.gene.list, xxx, sep = "\t")
cat(cmd12, file = file.path(temp3, paste0(xxx, "_config_cJunAndp27.txt")), sep = "\t")
}, cellInfo, temp3)
# dir.name=temp3 dir.name=reformatPath(dir.name)
file.name <-
list.files(
temp3,
pattern = "*_config_cJunAndp27.txt",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE
)
file.name.2 <- as.list(file.name)
zzz <- unlist(file.name.2)
lapply(1:length(zzz), function(u, zzz)
{
dir.name = dirname(zzz[u][[1]])
file_name = file_path_sans_ext(basename(zzz[u][[1]]))
cmd = paste(
"ngs.plot.r -G hg19 -R tss -C",
zzz[u][[1]],
"-O",
file.path(dir.name,
paste0(file_name, ".tss")),
"-T",
file_name,
"-L 4000 -RR 1 -CD 1 -GO total",
sep = " "
)
cmd2 = cmd
cat(cmd2, "\n")
cat("\n")
system(cmd2)
}, zzz)
return(re)
}
#mcf.with.zhao <- ChipSeq:::addMoreBed2mcf("/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny_zhao.csv","~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny","/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall","bed$",mcf=NULL,"/Volumes/Bioinformatics$/2017/DannyNewData/testRun5")
addMoreBed2mcf<-function(input.sample.file,input.bam.dir,input.bed.dir,input.file.pattern,mcf=NULL,output.dir){
bam.file <- prepareBamFile(input.bam.dir)
sample.info<-read.csv(input.sample.file,header = TRUE)
sample.info.2 <- sample.info[,-dim(sample.info)[2]]
#print(sample.info.2)
#out.dir.name = dirname(input.sample.file)
re<-ParserReadFiles(input.bed.dir,input.file.pattern)
re.bed<-re$input
#print(re.bed)
re.peaks.only.bed.2<-Filter(function(x) !any(grepl("summits", x)), re.bed)
# if(!is.null(summit_peak)){
# re.summits.only.bed<-FL(re.bed,'summits')
# }
#put peak files into database
re.peaks.only.bed.3<-list_to_df(re.peaks.only.bed.2)
print(re.peaks.only.bed.3)
ll <- re.peaks.only.bed.3$ID
ID2 <- unlist(lapply(ll,function(u) {
pos <- regexpr('R1',u)
if(pos>0){
id <- substr(u,1,pos+1)
}else{
id <- u
}
id
}))
# if(u>0){
# ID2 <- substr(ll,1,pos+1)
# }else{
# ID2 <- ll
# }
# pos <- regexpr('R1', re.peaks.only.bed.3$ID)
#
# print(pos)
#
# ll <- re.peaks.only.bed.3$ID
#
# ID2 <- lapply(pos,function(u,ll) {
#
# if(u>0){
# ID2 <- substr(ll,1,pos+1)
# }else{
# ID2 <- ll
# }
#
# },ll)
re.peaks.only.bed.3 <- cbind(ID2,re.peaks.only.bed.3)
#print(re.peaks.only.bed.3)
colnames(re.peaks.only.bed.3)=c("ID","ID2")
#
re.peaks.only.bed.4<-merge(sample.info.2,re.peaks.only.bed.3,by="ID")
#
cat("4\n")
print(re.peaks.only.bed.4)
cat("\n")
re.peaks.only.bed.with.bam<-merge(bam.file,re.peaks.only.bed.4,by="ID")
print(re.peaks.only.bed.with.bam)
colnames(re.peaks.only.bed.with.bam)[6]="SampleID"
replicate<-apply(re.peaks.only.bed.4,1,function(x){
# pos <- regexpr('R1', re.peaks.only.bed.3$ID)
ID=as.character(x[1])
pos <- regexpr('R1',ID)
if(pos>0){
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
p1=nchar(ID)+2
p2=regexpr(TF,ID2)
p2=p2-2
xx=substr(ID2,p1,p2)
cell=substr(xx,1,nchar(xx)-2)
rep=substr(xx,nchar(xx),nchar(xx))
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
}else
{
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
#p1=nchar(ID)+2
#p2=regexpr(TF,ID2)
#p2=p2-2
#xx=substr(ID2,p1,p2)
#cell=substr(xx,1,nchar(xx)-2)
rep=1
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
}
z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
z
})
res <- do.call(rbind.data.frame,replicate)
colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
colnames(res)[1] = "SampleID"
sample.4.dba<-merge(res,re.peaks.only.bed.with.bam,by="SampleID")
re.peaks.only.bed.5 <- res
# replicate<-apply(re.peaks.only.bed.4,1,function(x){
# ID=as.character(x[1])
# cell=as.character(x[2])
# TF=as.character(x[3])
# ID2=as.character(x[4])
# p1=nchar(ID)+2
# p2=regexpr(TF,ID2)
# p2=p2-2
# xx=substr(ID2,p1,p2)
# cell=substr(xx,1,nchar(xx)-2)
# #rep=1
# file=as.character(x[6])
# condition=paste(cell,TF,sep=":")
# z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
# z
# })
#
# res <- do.call(rbind.data.frame,replicate)
# colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
#
#
# re.peaks.only.bed.5 <- res
print(re.peaks.only.bed.5)
mcf<-mcf
ll <- list()
for(i in 1:dim(re.peaks.only.bed.5)[1]){
#peaks.v=readBed(as.character(re.peaks.only.bed.5[i,6]))
peaks.v <- readPeakFile(as.character(re.peaks.only.bed.5[i,6]), as = "GRanges")
peaks.v.sorted <- DiffBind:::pv.peaksort(as.data.frame(peaks.v))
#peaks.v.sorted <- peaks.v
##print(peaks.v)
ll[[i]] <- peaks.v.sorted
peak.caller.v="macs142"
sampID.v=as.character(re.peaks.only.bed.5[i,1])
pos <- regexpr('R1',sampID.v)
if(pos>0){
sampID.v=substr(sampID.v,1,nchar(sampID.v)-5)
}else{
sampID.v = sampID.v
}
tissue.v=as.character(re.peaks.only.bed.5[i,2])
factor.v=as.character(re.peaks.only.bed.5[i,3])
condition.v=as.character(re.peaks.only.bed.5[i,4])
replicate.v=as.numeric(as.character(re.peaks.only.bed.5[i,5]))
if(is.null(mcf)){
mcf <- dba.peakset(NULL,peaks=peaks.v.sorted,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}else{
mcf <- dba.peakset(mcf,peaks=peaks.v.sorted,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}
}
print(mcf)
temp3=output.dir
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
save(mcf,file=file.path(temp3,"mcf.RData"))
pdf(file.path(temp3,"corrmap.pdf"))
dba.plotHeatmap(mcf,margin=23)
dev.off()
# #GetResultsFromDiffBind(mcf,"yes",temp3)
re <- list(mcf=mcf,ll = ll,re.peaks.only.bed.5 = re.peaks.only.bed.5,re.peaks.only.bed.with.bam = re.peaks.only.bed.with.bam, sample.4.dba = sample.4.dba)
return(re)
}
#ChipSeq:::GetResultsFromDiffBind2(temp,"/Volumes/Bioinformatics$/2017/DannyNewData/testRun2")
GetResultsFromDiffBind2<-function(mcf7,output.file.dir){
temp<-mcf.with.zhao
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
#if(Mergereplicates=="yes"){
temp22<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp22)
##need to set the flexiable number identified from data sets
# t <- temp2
# temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
# }else{
# temp22<-temp
# }
temp22<-dba(temp22,mask =17:28)
print(temp22)
po1 <- c(1,2,6,10)
po2 <- c(1,2)
temp3=file.path(output.file.dir,"Venn2")
if(!dir.exists(temp3))
{
dir.create(temp3)
}
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po1,main=paste0(colnames(temp22$class)[po1],collapse = "-vs-"))
dev.off()
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po2,main=paste0(colnames(temp22$class)[po2],collapse = "-vs-"))
dev.off()
# Tissue1<-temp22$class[2,]
# Tissue2<-unique(temp22$class[2,])
#
# TF<-unique(temp22$class[3,])
# TF.n<-length(TF)
#
# temp3=file.path(output.file.dir,"Venn")
#
# if(!dir.exists(temp3))
# {
# dir.create(temp3)
# }
#
# for(i in 1:length(Tissue2)){
# po<-which(Tissue1 %in% Tissue2[i])
#
# print(po)
#
# if(length(po)==2)
# {
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po,main=Tissue2[i])
# dev.off()
#
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
# dev.off()
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
# dev.off()
#
# }else
# {
# cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
# }
# }
#
# p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
#
# po<-which(Tissue1 %in% Tissue2[u])
#
# if(length(po)==2)
# {
# common.peaks<-dba.overlap(temp22,mask=po)
# y<-common.peaks$inAll
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# common.peaks.1<-dba.overlap(temp22,mask=po1)
# y1<-common.peaks.1$inAll
#
# common.peaks.2<-dba.overlap(temp22,mask=po2)
# y2<-common.peaks.2$inAll
#
# y<-list(y1=y1,y2=y2)
#
# names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
# names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
#
# }else{
# y<-NULL}
# y
# },Tissue1,Tissue2,temp22)
#
# names(p.common)<-Tissue2
#
# p.common<-unlist(p.common,recursive = F)
#
# p.common<-p.common[lapply(p.common,length) > 0]
#
# if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
# {
# dir.create(file.path(output.file.dir,"common_peaks_bed"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
# }
#
# #output common peaks to bed files
#
# lapply(1:length(p.common),function(u,p.common,output.file.dir){
# x=p.common[[u]]
#
# x_name=names(p.common)[u]
#
# df <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=strand(x))
#
# #assign strand
# df.str <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=c(rep(".", length(x))))
#
# df.str.1<-df.str[-grep("random",df.str$seqnames),]
#
# df.str.2<-df.str.1
#
# df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
# col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# },p.common,output.file.dir)
#
# AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
# ,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
#
# return(p.common)
}
useChIPpeakAnno <- function(mcf.with.zhao) {
#findOverlapsOfPeaks(mcf.with.zhao$ll[c(17,11,18)])
overlap <- findOverlapsOfPeaks(mcf.with.zhao$ll[17][[1]],mcf.with.zhao$ll[11][[1]],mcf.with.zhao$ll[18][[1]])
bed.li<- overlap$overlappingPeaks[2][[1]][,2:4]
write.table(bed.li,file="cJun_Li.bed",sep="\t",row.names = FALSE,
col.names = FALSE,quote=FALSE)
bed.zhao <- overlap$overlappingPeaks[2][[1]][,8:10]
write.table(bed.zhao,file="cJun_zhao.bed",sep="\t",row.names = FALSE,
col.names = FALSE,quote=FALSE)
s1 <- makeVennDiagram(mcf.with.zhao$ll[c(17,11,18)], NameOfPeaks=c("cJun_Li","cJun_1833","cJun_zhao"),
totalTest=20000,scaled=F, euler.d=F,fill = c("red","green","blue"),
alpha = 0.50,
label.col = c(rep("black",7)),
cex = 2,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","green","blue"),connectedPeaks = "keepAll")
s2 <- makeVennDiagram(mcf.with.zhao$ll[c(17,3,18)], NameOfPeaks=c("cJun_Li","cJun_231","cJun_zhao"),
totalTest=20000,scaled=F, euler.d=F,fill = c("red","green","blue"),
alpha = 0.50,
label.col = c(rep("black",7)),
cex = 2,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","green","blue"),connectedPeaks = "keepAll")
}
# SampleID,Tissue,Factor,Condition,Treatment,Replicate,bamReads,ControlID,bamControl,Peaks,PeakCaller
# BT4741,BT474,ER,Resistant,Full-Media,1,reads/Chr18_BT474_ER_1.bam,BT474c,reads/Chr18_BT474_input.bam,peaks/BT474_ER_1.bed.gz,bed
# BT4742,BT474,ER,Resistant,Full-Media,2,reads/Chr18_BT474_ER_2.bam,BT474c,reads/Chr18_BT474_input.bam,peaks/BT474_ER_2.bed.gz,bed
# MCF71,MCF7,ER,Responsive,Full-Media,1,reads/Chr18_MCF7_ER_1.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_1.bed.gz,bed
# MCF72,MCF7,ER,Responsive,Full-Media,2,reads/Chr18_MCF7_ER_2.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_2.bed.gz,bed
# MCF73,MCF7,ER,Responsive,Full-Media,3,reads/Chr18_MCF7_ER_3.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_3.bed.gz,bed
# T47D1,T47D,ER,Responsive,Full-Media,1,reads/Chr18_T47D_ER_1.bam,T47Dc,reads/Chr18_T47D_input.bam,peaks/T47D_ER_1.bed.gz,bed
# T47D2,T47D,ER,Responsive,Full-Media,2,reads/Chr18_T47D_ER_2.bam,T47Dc,reads/Chr18_T47D_input.bam,peaks/T47D_ER_2.bed.gz,bed
# MCF7r1,MCF7,ER,Resistant,Full-Media,1,reads/Chr18_TAMR_ER_1.bam,TAMRc,reads/Chr18_TAMR_input.bam,peaks/TAMR_ER_1.bed.gz,bed
# MCF7r2,MCF7,ER,Resistant,Full-Media,2,reads/Chr18_TAMR_ER_2.bam,TAMRc,reads/Chr18_TAMR_input.bam,peaks/TAMR_ER_2.bed.gz,bed
# ZR751,ZR75,ER,Responsive,Full-Media,1,reads/Chr18_ZR75_ER_1.bam,ZR75c,reads/Chr18_ZR75_input.bam,peaks/ZR75_ER_1.bed.gz,bed
# ZR752,ZR75,ER,Responsive,Full-Media,2,reads/Chr18_ZR75_ER_2.bam,ZR75c,reads/Chr18_ZR75_input.bam,peaks/ZR75_ER_2.bed.gz,bed
#prepareBamFile("~/Dropbox (BBSR)/BWA")
prepareBamFile <- function(input.bam.dir,file.pattern){
bam <- list.files(path = input.bam.dir, pattern=file.pattern, all.files = TRUE,
full.names = TRUE, recursive = TRUE,
ignore.case = FALSE, include.dirs = TRUE, no.. = FALSE)
bam2 <- as.data.frame(cbind(basename(bam),bam))
ID <- unlist(lapply(bam2[,1],function(u)
{
y <- u
pos <- regexpr("\\.", y)
pos <- pos - 1
y <- substr(y, 1, pos)
y
}))
bam3 <- cbind(ID,bam2)
bam3
}
# tamoxifen <- prepareSampleSheet(mcf.with.zhao,"~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny")
prepareSampleSheet <- function(mcf.with.zhao,output.file.dir){
sampleSheet <- mcf.with.zhao$sample.4.dba[,c(1,2,3,5,9,13)]
sampleSheet2 <- cbind(sampleSheet[,1:3],rep("Danny-Treatment",dim(sampleSheet)[1]),rep("Danny-condition",dim(sampleSheet)[1]),sampleSheet[,4:6],rep("bed",dim(sampleSheet)[1]))
colnames(sampleSheet2)=c("SampleID","Tissue","Factor","Condition","Treatment","Replicate","bamReads","Peaks","PeakCaller")
fwrite(sampleSheet2, file ="sampleSheet2.csv")
tamoxifen <- dba(sampleSheet="sampleSheet2.csv")
tamoxifen
tamoxifen <- dba.count(tamoxifen, summits=250)
tamoxifen <- dba.contrast(tamoxifen, group1 = c(1,2),group2=c(3,4),name1="cJun_231_DD", name2="cJun_231")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(1,2),group2=c(11,12),name1="cJun_231_DD", name2="cJun_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(3,4),group2=c(11,12),name1="cJun_231", name2="cJun_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(13,14),group2=c(5,6),name1="p27_231_DD", name2="p27_231")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(13,14),group2=c(15,16),name1="p27_231_DD", name2="p27_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(5,6),group2=c(15,16),name1="p27_231", name2="p27_1833")
tamoxifen <- dba.analyze(tamoxifen)
save(tamoxifen,file=file.path(output.file.dir,"Danny_Dba.RData"))
tamoxifen
# dba.report(tamoxifen,contrast = 1)
# dba.report(tamoxifen,contrast = 2)
# dba.report(tamoxifen,contrast = 3)
# dba.report(tamoxifen,contrast = 4)
# dba.report(tamoxifen,contrast = 5)
# dba.report(tamoxifen,contrast = 6)
}
#generateBedFromDba(tamoxifen,"~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed")
generateBedFromDba <- function(tamoxifen,output.file.dir){
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
n <- length(tamoxifen$contrasts)
lapply(1:n,function(u,tamoxifen){
temp <- as.data.frame(dba.report(tamoxifen,contrast = u,th=1))
file.name <- paste(colnames(as.data.frame(dba.report(tamoxifen,contrast = u,th=1)))[c(7,8)],collapse = "-")
write.table(temp, row.names=F, col.names = F, quote =F,sep="\t", file=file.path(output.file.dir,paste0(file.name,".bed")))
},tamoxifen)
}
#' dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed"
#' input.file.pattern=".bed"
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnntationUsingChipSeeker2 <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=5000,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
re<-ParserReadFiles(dir.name,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2 <- re.bed
# if(length(dir(dir.name,pattern="peaks.bed"))!=0)
# {
# re.peaks.only.bed.2<-FL(re.bed,'peaks')
# cat("peaks\n")
# print(re.peaks.only.bed.2)
# }
#
# if(length(dir(dir.name,pattern="summits.bed"))!=0){
# re.summits.only.bed<-FL(re.bed,'summits')
# cat("summits\n")
# print(re.summits.only.bed)
# }
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,"Annotation",APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
peaks.anno.list <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
#select.index <- which(peakAnno$distanceToTSS<20,000 && peakAnno$distanceToTSS >= -20,000)
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
x_name=names(re.peaks.only.bed.2)[u]
cat(x_name,"\n")
png(file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_pie.png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
#print(paste0(peaks[,c(2,3)]))
group1 <- strsplit(tools::file_path_sans_ext(x_name),"-")[[1]][1]
group2 <- strsplit(tools::file_path_sans_ext(x_name),"-")[[1]][2]
colnames(peaks.anno)[5:13]=c("starnd","width_DiffDind_based","strand","Conc", group1,group2,"Fold","p.value","FDR")
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_only_mapped_peaks.xls")),
row.names = FALSE,quote=FALSE,sep="\t")
# unmapped.peaks<-peaks[-which(paste0(peaks[,2],"_",peaks[,3]) %in% paste0(peaks.anno[,2],"_",peaks.anno[,3])),]
#
# cat(dim(peaks)[1]," ",dim(peaks.anno)[1]," ",dim(unmapped.peaks)[1],"\n")
#
#
# if(dim(unmapped.peaks)[1]!=0){
#
# colnames(unmapped.peaks)=colnames(peaks.anno)[1:6]
#
# unmapped.peaks.3<-smartbind(peaks.anno,unmapped.peaks)
#
# unmapped.peaks.4<-unmapped.peaks.3[order(unmapped.peaks.3[,1],unmapped.peaks.3[,2]),]
#
# write.table(unmapped.peaks.4,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_all_peaks.xls")),row.names = FALSE,quote=FALSE,sep="\t")
# }
#
peaks.anno
},re.peaks.only.bed.2,d)
}
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/CombineChipSeqWithRNASeq"
#' rna.seq.input.file="/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx"
#'
#' integrateChipSeqWithRNASeq(out.dir.name,rna.seq.input.file)
#'
integrateChipSeqWithRNASeq <- function(output.file.dir,rna.seq.input.file) {
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
gene.231dd.231.1833.1833shp27 <- read.xlsx(rna.seq.input.file,sheetIndex = 1,header = FALSE)
colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
data.231DD.231 <- gene.231dd.231.1833.1833shp27[,c(1,2,3)]
data.1833.231 <- gene.231dd.231.1833.1833shp27[,c(1,4,5)]
data.1833shp27.1833 <- gene.231dd.231.1833.1833shp27[c(1,6,7)]
cJun.231DD.231.ChipSeq.RNASeq <- merge(res.promoter[[2]],data.231DD.231,by="SYMBOL")
p27.231DD.231.ChipSeq.RNASeq <- merge(res.promoter[[5]],data.231DD.231,by="SYMBOL")
cJun.231.1833.ChipSeq.RNASeq <- merge(res.promoter[[3]],data.1833.231,by="SYMBOL")
p27.231.1833.ChipSeq.RNASeq <- merge(res.promoter[[6]],data.1833.231,by="SYMBOL")
write.table(cJun.231DD.231.ChipSeq.RNASeq,file=file.path(output.file.dir,"cJun_231DD_231_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(p27.231DD.231.ChipSeq.RNASeq,file=file.path(output.file.dir,"p27_231DD_231_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(cJun.231.1833.ChipSeq.RNASeq,file=file.path(output.file.dir,"cJun_231_1833_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(p27.231.1833.ChipSeq.RNASeq,file=file.path(output.file.dir,"p27_231_1833_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
}
addTestFunction4HeatmapChipSeq <- function(GenomicRanges, rtracklayer, IRanges, dataDirectory, chipseq, VennDiagram) {
library(GenomicRanges)
library(rtracklayer)
library(IRanges)
input = import.bed(file.path(dataDirectory, 'ES_input_filtered_ucsc_chr6.bed'),
asRangedData=FALSE)
rep1 = import.bed("/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/2017-03-02-01_S5_R1-MDA-MB-231-DD-1-cJun_hs_1.00e-05_macs142_peaks.bed",genome = "hg19")
rep2 = import.bed("/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/2017-03-02-02_S2_R1-MDA-MB-231-DD-2-cJun_hs_1.00e-05_macs142_peaks.bed",genome = "hg19")
library(chipseq)
estimate.mean.fraglen(rep1)
prepareChIPseq = function(reads){
frag.len = median( estimate.mean.fraglen(reads) )
cat( paste0( 'Median fragment size for this library is ', round(frag.len)))
reads.extended = resize(reads, width = frag.len)
return( trim(reads.extended) )
}
rep1 = prepareChIPseq(rep1)
ovlp = findOverlaps(rep1,rep2)
ovlp = as.data.frame(ovlp)
ov = min( length(unique(ovlp$queryHits)), length(unique(ovlp$subjectHits)) )
library(VennDiagram)
draw.pairwise.venn(
area1=length(rep1),
area2=length(rep2),
cross.area=ov,
category=c("rep1", "rep2"),
fill=c("steelblue", "blue3"),
cat.cex=0.7)
}
#input.region.bed.dir = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
#plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin","hg19_gene.bed",)
#
plotHeatMapUsedeepTools <- function(input.sample.file,input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
bw.file.sample.label <- mapBw3Sample(input.sample.file,input.bw.file.dir)
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
bed.file.list <- list.files(
input.region.bed.dir,
pattern = "*.bed$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
bw.file.list <- as.character(bw.file.sample.label$file.bw)
samplesLabel <- as.character(bw.file.sample.label$sampleLabel)
print(bed.file.list)
if(select.region.bed!=""){
input.beds = bed.file.list[-grep(paste(select.region.bed,collapse = "|"), bed.file.list)]
input.beds = paste(input.beds,collapse = " ")
}else{
input.beds = paste(bed.file.list,collapse = " ")
}
print(input.beds)
cmd = "computeMatrix reference-point --referencePoint TSS -b 4000 -a 4000 -R"
input.beds=input.beds
input.bw.file=paste(bw.file.list,collapse = " ")
input.samplesLabel=paste(samplesLabel,collapse = " ")
outFileNameMatrix=file.path(output.file.dir,"matrix1_cJun_p27_TSS.gz")
outFileSortedRegions=file.path(output.file.dir,"regions_cJun_p27_genes.bed")
outHeatMapFile=file.path(output.file.dir,"heatmap_cJun_p27.png")
cmd1=paste(cmd,input.beds,"-S",input.bw.file,"--skipZero","-o",outFileNameMatrix,"--outFileSortedRegions",outFileSortedRegions,"--samplesLabel",input.samplesLabel,collapse = " ")
cmd2=paste("plotHeatmap -m",outFileNameMatrix,"-out",outHeatMapFile,collapse = " ")
#
# testFiles/genes.bed -S testFiles/log2ratio_H3K4Me3_chr19.bw --skipZeros
# -o matrix1_H3K4me3_l2r_TSS.gz
# --outFileSortedRegions regions1_H3K4me3_l2r_genes.bed
#
# deepTools2.0/bin/computeMatrix scale-regions \
# -R genes_chr19_firstHalf.bed genes_chr19_secondHalf.bed \ # separate multiple files with spaces
# -S testFiles/log2ratio_*.bw \ or use the wild card approach
# -b 3000 -a 3000 \
# --regionBodyLength 5000 \
# --skipZeros -o matrix2_multipleBW_l2r_twoGroups_scaled.gz \
# --outFileNameMatrix matrix2_multipleBW_l2r_twoGroups_scaled.tab \
# --outFileSortedRegions regions2_multipleBW_l2r_twoGroups_genes.bed
#
# computeMatrix reference-point \ # choose the mode
# --referencePoint TSS \ # alternatives: TES, center
# -b 3000 -a 10000 \ # define the region you are interested in
# -R testFiles/genes.bed \
# -S testFiles/log2ratio_H3K4Me3_chr19.bw \
# --skipZeros \
# -o matrix1_H3K4me3_l2r_TSS.gz \ # to be used with plotHeatmap and plotProfile
# --outFileSortedRegions regions1_H3K4me3_l2r_genes.bed
#cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd1=paste(cmd.java.2,cmd1,sep=";")
system(cmd1)
#print(cmd2)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin/annotation","hg19_gene.bed","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4plotHeatMapUsedeepTools <- function(input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "HeapMapPlot"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::plotHeatMapUsedeepTools('
Rinput <- paste0('\\"',input.bw.file.dir,'\\",',
'\\"',input.region.bed.dir,'\\",',
'\\"',select.region.bed,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
}
# Welcome to Rattler, *please* read these important system notes:
#
# --> Rattler is currently running the SLURM resource manager to
# schedule all compute resources. Example SLURM job scripts are
# available on the system at /cm/shared/docs/slurm/current/job_scripts
#
# To run an interactive shell, issue:
# srun -p hipri -t 0:30:00 -n 36 -N 1 --pty /bin/bash -l
#
# To submit a batch job, issue: sbatch job_script
# To show all queued jobs, issue: squeue
# To kill a queued job, issue: scancel <jobId>
#
# See "man slurm" for more detailed information.
#
# --> Rattler has 3 main queues with the following timelimits:
# * hipri (2 days)
# * iq (1 day)
# * requestq (infinite) - Requires a valid slurm reservation to use
#
# --> To see which software packages are available issue: "module avail"
#
# Use the following commands to adjust your environment:
# 'module add <module>' - adds a module to your environment for this session
# 'module initadd <module>' - configure module to be loaded at every login
#
#plotHeatMapUsedeepTools("/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap/matrix1_cJun_p27_TSS.gz","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")
#
usePlotHeatmap <- function(input.matrix.file,output.file.dir){
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
outHeatMapFile=file.path(output.file.dir,"heatmap_cJun_p27.png")
cmd2=paste("plotHeatmap -m",input.matrix.file,"-out",outHeatMapFile,collapse = " ")
system(cmd2)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4usePlotHeatmap("/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap/matrix1_cJun_p27_TSS.gz","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4usePlotHeatmap <- function(input.matrix.file,output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "plotHeatmap"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::usePlotHeatmap('
Rinput <- paste0('\\"',input.matrix.file,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
system(cmd.gff)
}
# input.bam.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed/Annotation"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
# generateBed4HeatMap(input.bam.file.dir,out.dir.name)
#
generateBed4HeatMap <- function(input.bam.file.dir,out.dir.name) {
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.list <- list.files(
input.bam.file.dir,
pattern = "*xls$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
file.table <- lapply(file.list,function(u){
y <- basename(u)
pos <- regexpr("\\_", y)
pos <- pos - 1
y <- substr(y, 1, pos)
x <-fread(u)
x<- cbind(x,rep(y,dim(x)[1]))
colnames(x)[dim(x)[2]]="sample_name"
x
})
# library(dplyr)
# x <- data_frame(i = c("a","b","c"), j = 1:3)
# y <- data_frame(i = c("b","c","d"), k = 4:6)
# z <- data_frame(i = c("c","d","a"), l = 7:9)
common.gene.peak <- file.table %>%
Reduce(function(dtf1,dtf2) inner_join(dtf1,dtf2,by="SYMBOL"),.)
x <- data.table::subset(common.gene.peak,select=c("geneChr.x","geneStart.x","geneEnd.x","geneStrand.x","SYMBOL"))
data.table::setkey(x,NULL)
xx <- unique(x)
xxx<-xx[complete.cases(xx),]
# gene.231dd.231.1833.1833shp27 <- read.xlsx(
# "/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx",
# sheetIndex = 1,header = FALSE)
# colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
#save(gene.231dd.231.1833.1833shp27,file="./data/gene.RData")
gene.45 <- merge(gene.231dd.231.1833.1833shp27,xxx,by="SYMBOL",sort = FALSE)
gene.45.1 <- gene.45[,c(8,9,10,11,1)]
colnames(gene.45.1)=c("chr","start","end","strand","SYMBOL")
gene.45.1[which(gene.45.1$strand==1),]$strand="+"
gene.45.1[which(gene.45.1$strand==2),]$strand="-"
gene.45.1.sorted.by.chr <- gene.45.1[order(gene.45.1$chr),]
gene.45.1$chr<-paste0("chr",gene.45.1$chr)
gene.45.1.sorted.by.chr$chr<-paste0("chr",gene.45.1.sorted.by.chr$chr)
write.table(gene.45.1,file=file.path(out.dir.name,"gene_200.bed"),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
write.table(gene.45.1.sorted.by.chr,file=file.path(out.dir.name,"gene_200_sorted.bed"),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("~/SampleID_INFO_ChIP_new_Danny.csv","~/BamCompare","~/ChipSeqBed",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmapAddlabel")'
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("~/SampleID_INFO_ChIP_new_Danny.csv","~/BamCoverage","~/ChipSeqBed_p27_cJun",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap4p27_cJun")'
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("/nethome/axy148/R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","~/BamCompareZhao","~/cJun_gene_bed",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/zhao_ChipSeq_heatmap4cJun")'
bashJob4plotHeatMapUsedeepTools <- function(input.sample.file,input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "generateMatrix"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::plotHeatMapUsedeepTools('
Rinput <- paste0('\\"',input.sample.file,'\\",',
'\\"',input.bw.file.dir,'\\",',
'\\"',input.region.bed.dir,'\\",',
'\\"',select.region.bed,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
job.name <- "plotHeatmap"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::usePlotHeatmap('
input.matrix.file <- file.path(output.file.dir,"matrix1_cJun_p27_TSS.gz")
Rinput <- paste0('\\"',input.matrix.file,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name="generateMatrix")
system(cmd.gff)
}
#input.bw.dir="~/BamCompare"
#input.sample.file="~/SampleID_INFO_ChIP_new_Danny.csv"
#
#mapBw3Sample(input.sample.file,input.bw.dir)
#
mapBw3Sample <- function(input.sample.file,input.bw.dir) {
file.1 <- list.files(input.bw.dir,pattern=".bw$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p1 <- regexpr("\\_", x);p2 <- regexpr("\\.", x);xx <- substr(x,p1+1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bw")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
makeHeatMapByme <- function() {
compute.matrix.file="/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Danny_ChipSeq_heatmapBasedSortedBed/matrix1_cJun_p27_TSS.gz"
table = read.table(gzfile(compute.matrix.file), skip=1)
matrix = table[7:dim(table)[2]]
image(1:dim(matrix)[2], 1:dim(matrix)[1], t(as.matrix(matrix)), axes=FALSE, xlab='sample', ylab='gene')
axis(2, at=1:length(table$V1), labels=table$V4, las = 1)
}
# input.bam.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed/Annotation"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name)
#
# input.bam.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed/p27"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name,"p27")
#
# input.bam.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed/cJun"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name,"cJun")
#
generateBed4HeatMap2 <- function(input.bam.file.dir,out.dir.name,Ab) {
out.dir.name <-file.path(out.dir.name,paste0(Ab,"_gene_bed"))
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.list <- list.files(
input.bam.file.dir,
pattern = "*xls$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
file.table <- lapply(file.list,function(u){
y <- basename(u)
pos <- regexpr("\\_", y)
pos <- pos - 1
y <- substr(y, 1, pos)
x <-fread(u)
x<- cbind(x,rep(y,dim(x)[1]))
colnames(x)[dim(x)[2]]="sample_name"
x
})
# library(dplyr)
# x <- data_frame(i = c("a","b","c"), j = 1:3)
# y <- data_frame(i = c("b","c","d"), k = 4:6)
# z <- data_frame(i = c("c","d","a"), l = 7:9)
common.gene.peak <- file.table %>%
Reduce(function(dtf1,dtf2) full_join(dtf1,dtf2,by="SYMBOL"),.)
x <- subset(common.gene.peak,select=c("geneChr.x","geneStart.x","geneEnd.x","geneStrand.x","SYMBOL"))
setkey(x,NULL)
xx <- unique(x)
xxx<-xx[complete.cases(xx),]
# gene.231dd.231.1833.1833shp27 <- read.xlsx(
# "/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx",
# sheetIndex = 1,header = FALSE)
# colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
#save(gene.231dd.231.1833.1833shp27,file="./data/gene.RData")
#gene.45 <- merge(gene.231dd.231.1833.1833shp27,xxx,by="SYMBOL",sort = FALSE)
gene.45 <- xxx
gene.45.1 <- gene.45
colnames(gene.45.1)=c("chr","start","end","strand","SYMBOL")
gene.45.1[which(gene.45.1$strand==1),]$strand="+"
gene.45.1[which(gene.45.1$strand==2),]$strand="-"
gene.45.1.sorted.by.chr <- gene.45.1[order(gene.45.1$chr),]
gene.45.1$chr<-paste0("chr",gene.45.1$chr)
gene.45.1.sorted.by.chr$chr<-paste0("chr",gene.45.1.sorted.by.chr$chr)
write.table(gene.45.1,file=file.path(out.dir.name,paste0("gene_",Ab,".bed")),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
write.table(gene.45.1.sorted.by.chr,file=file.path(out.dir.name,paste0("gene_",Ab,"_sorted.bed")),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
# > length(unique(file.table[[1]]$SYMBOL))
# [1] 417
# > length(unique(file.table[[2]]$SYMBOL))
# [1] 222
# > length(unique(file.table[[3]]$SYMBOL))
# [1] 458
#
#
# unique(c(unique(file.table[[1]]$SYMBOL),unique(file.table[[2]]$SYMBOL),unique(file.table[[3]]$SYMBOL)))
gene.45.1.sorted.by.chr
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "BamCoverage" -o %J.BamCoverage.log -e %J.BamCoverage.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCoverage("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/nethome/axy148/_visualization/sorted_bam_files_2.txt","~/BamCoverage")'
useBamCoverage <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
#bamCoverage -b reads.bam -o coverage.bw
#bamCoverage --bam a.bam -o a.SeqDepthNorm.bw \
#--binSize 10
#--normalizeTo1x 2150570000
#--ignoreForNormalization chrX
#--extendReads
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX"
cmd3 <- "-o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("To1x_",basename(as.character(xx)),"_vs_","no_input",".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
#' output.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
#' #mcf7=mcf2
#' re<-getTargetGene4Ab(mcf2,"yes","p27",output.file.dir)
#' re<-getTargetGene4Ab(mcf2,"yes","cJun",output.file.dir)
#'
getTargetGene4Ab<-function(mcf7,Mergereplicates=c("yes","no"),Ab,output.file.dir){
output.file.dir = file.path(output.file.dir,Ab)
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir,recursive = TRUE)
}
temp<-mcf7
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
if(Mergereplicates=="yes"){
temp2<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp2)
##need to set the flexiable number identified from data sets
t <- temp2
temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
}else{
temp22<-temp
}
print(temp22)
#temp22 <- re
Tissue1<-temp22$class[2,]
Tissue2<-unique(temp22$class[2,])
TF<-unique(temp22$class[3,])
TF.n<-length(TF)
for(i in 1:length(Tissue2)){
#for(j in 1:length(TF)){
p <- which(temp22$class[2,]==Tissue2[i]&temp22$class[3,]==Ab)
cat(Tissue2[i]," ",Ab," ",p,"\n")
if(length(p)!=0){
#as.data.frame(dba.peakset(temp22,peaks=1,bRetrieve=TRUE))
dba.peakset(temp22,peaks=as.integer(p),bRetrieve=TRUE, writeFile = file.path(output.file.dir,paste0(Tissue2[i],"_",Ab,".bed")))
}
}
AnntationUsingChipSeeker3(output.file.dir,".bed",output.file.dir
,txdb="hg19",DD=5000)
# AnntationUsingChipSeeker2
# temp3=file.path(output.file.dir,"Venn")
#
# if(!dir.exists(temp3))
# {
# dir.create(temp3)
# }
#
# for(i in 1:length(Tissue2)){
# po<-which(Tissue1 %in% Tissue2[i])
#
# print(po)
#
# if(length(po)==2)
# {
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po,main=Tissue2[i])
# dev.off()
#
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
# dev.off()
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
# dev.off()
#
# }else
# {
# cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
# }
# }
#
# p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
#
# po<-which(Tissue1 %in% Tissue2[u])
#
# if(length(po)==2)
# {
# common.peaks<-dba.overlap(temp22,mask=po)
# y<-common.peaks$inAll
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# common.peaks.1<-dba.overlap(temp22,mask=po1)
# y1<-common.peaks.1$inAll
#
# common.peaks.2<-dba.overlap(temp22,mask=po2)
# y2<-common.peaks.2$inAll
#
# y<-list(y1=y1,y2=y2)
#
# names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
# names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
#
# }else{
# y<-NULL}
# y
# },Tissue1,Tissue2,temp22)
#
# names(p.common)<-Tissue2
#
# p.common<-unlist(p.common,recursive = F)
#
# p.common<-p.common[lapply(p.common,length) > 0]
#
# if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
# {
# dir.create(file.path(output.file.dir,"common_peaks_bed"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
# }
#
# #output common peaks to bed files
#
# lapply(1:length(p.common),function(u,p.common,output.file.dir){
# x=p.common[[u]]
#
# x_name=names(p.common)[u]
#
# df <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=strand(x))
#
# #assign strand
# df.str <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=c(rep(".", length(x))))
#
# df.str.1<-df.str[-grep("random",df.str$seqnames),]
#
# df.str.2<-df.str.1
#
# df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
# col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# },p.common,output.file.dir)
#
# AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
# ,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
#
# return(p.common)
}
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("~/SampleID_INFO_ChIP_new_Danny.csv","~/sort_bam","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_data_QC")'
#
#R/lib64/R/library/ChipSeq/extdata/zhao_data.csv
#
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","~/sort_bam","/scratch/projects/bbc/aiminy_project/zhao_ChipSeq_data_QC")'
#
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("~/SampleID_INFO_ChIP_new_Danny.csv","/projects/scratch/bbc/Project/Danny_chip2/BigWig","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_data_QC_2")'
#
#
#
useRunSppR <- function(input.sample.file,input.bam.file.dir,output.file.dir){
#bam.file.sample.label <- mapBam2Sample2(input.sample.file,input.bam.file.dir)
bam.file.sample.label <- mapBam2Sample3(input.sample.file,input.bam.file.dir)
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
bam.file.list <- as.character(bam.file.sample.label$file.bam)
samplesLabel <- as.character(bam.file.sample.label$sampleLabel)
lapply(1:length(bam.file.list),function(u,bam.file.list,samplesLabel,output.file.dir){
cmd = paste("Rscript ~/phantompeakqualtools/run_spp.R",
paste0("-c=",bam.file.list[u]),
paste0("-savp=",file.path(output.file.dir,paste0(samplesLabel[u],"_QC.pdf"))),
paste0("-out=",file.path(output.file.dir,paste0(samplesLabel[u],"_QC_metrix.txt"))),sep=" ")
system(cmd)
},bam.file.list,samplesLabel,output.file.dir)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin/annotation","hg19_gene.bed","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4useRunSppR <- function(input.sample.file,input.bam.file.dir,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "QC"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::useRunSppR('
Rinput <- paste0('\\"',input.sample.file,'\\",',
'\\"',input.bam.file.dir,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- DoGs:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
system(cmd.gff)
}
#input.bw.dir="~/BamCompare"
#input.sample.file="~/SampleID_INFO_ChIP_new_Danny.csv"
#
#mapBw3Sample(input.sample.file,input.bw.dir)
#
mapBam2Sample <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p1 <- regexpr("\\_", x);p2 <- regexpr("\\.", x);xx <- substr(x,p1+1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
mapBam2Sample2 <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p2 <- regexpr("\\.", x);xx <- substr(x,1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
mapBam2Sample3 <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);xx <- tools::file_path_sans_ext(x)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
#' dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed"
#' input.file.pattern=".bed"
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnotationUsingChipSeeker3 <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=NULL,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
re<-ParserReadFiles(dir.name,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2 <- re.bed
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
peaks.anno.list <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
if(dim(peaks)[1]>0){
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
if(!is.null(distanceToTSS_cutoff)){
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
}else
{
peakAnno <- peakAnno
}
x_name=names(re.peaks.only.bed.2)[u]
x_name=tools::file_path_sans_ext(x_name)
x_name=tools::file_path_sans_ext(x_name)
cat(x_name,"\n")
png(file.path(temp3,paste0(x_name,"_",d,".png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,".xls")),
row.names = FALSE,quote=FALSE,sep="\t")
peaks.anno
}
},re.peaks.only.bed.2,d)
}
# R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4generateBed4HeatMap2("~/cJun","~/","cJun")'
bashJob4generateBed4HeatMap2 <- function(input.bam.file.dir,out.dir.name,Ab){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
job.name <- "generateGeneBed"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::generateBed4HeatMap2('
Rinput <- paste0('\\"',input.bam.file.dir,'\\",',
'\\"',out.dir.name,'\\",',
'\\"',Ab,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "bamCompare" -o %J.bamCompare.log -e %J.Compare.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCompare2("/nethome/axy148/R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","/nethome/axy148/_visualization/sort_bam.txt","~/BamCompareZhao")'
useBamCompare2 <- function(input.sample.file,input.bam.file,output.dir)
{
#sampleF <- read.table(input.sample.file,header = FALSE)
#colnames(sampleF) <- c("ID","SampleName")
#bam <- prepareBamFile(input.bam.file,file.pattern)
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1",xx,"--bamfile2",x.input,sep=" ")
cmd2 <- "--binSize 25"
cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(x.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
system("module unload python/2.7.3")
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::ngs()'
ngs <- function(){
system("ngs.plot.r -G hg19 -R tss -C ~/zhao_config.txt -O zhao.tss -L 4000")
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::ngs2()'
ngs2 <- function(config.file,distance.around.tss,output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
cmd=paste("ngs.plot.r -G hg19 -R tss -C",config.file,"-O",output.file.dir,"-L",distance.around.tss,"-CO white:white:red", collapse = " ")
system(cmd)
}
doAlignment<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
filterSam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd2='samtools view -Sh /home/someusername/myFastqFile.sam | grep -e "^@" -e "XM:i:[012][^0-9]" | grep -v "XS:i:" > /home/someusername/myFastqFile.sam.filtered.sam'
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
convertSam2Bam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd3="samtools view -S -b /home/someusername/myFastqFile.sam.filtered.sam > /home/someusername/myFastqFile.sam.filtered.sam.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
sortBam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd4="samtools sort /home/someusername/myFastqFile.sam.filtered.sam.bam /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
removePCRduplicates<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd5="samtools rmdup -s /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
indexBAM<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd6="samtools index /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
bam2Bed<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd7="bedtools bamtobed -i /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam > /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam.bed"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
getBamReady<-function(input.fastq.dir,bowtie2Indecesinput,sample.num,output.bam.dir){
Rfun1 <- 'library(ChipSeq);library(DoGs);re <- ChipSeq:::doAlignment('
Rinput <- paste0('\\"',input.fastq.dir,'\\",',
'\\"',bowtie2Indecesinput,'\\",',
'\\"',output.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
alignment <- createBsubJobArrayRfun(Rfun,paste0("alignment[1-",sample.num,"]",NULL))
system(alignment)
}
getEffectOfRmDup<-function(){
not.rm <- read.table("~/Dropbox (BBSR)/Aimin_project/Research/ChipSeq/Danny_ChipSeq_data_QC/qc_16.txt")
rm <- read.table("~/Dropbox (BBSR)/Aimin_project/Research/ChipSeq/Danny_ChipSeq_data_QC_2/16_samples_QC.txt")
res <- as.data.table(rbind(cbind(rep("not_rm",length(not.rm[,11])),not.rm[,11]),cbind(rep("rm",length(rm[,11])),rm[,11])))
boxplot(as.numeric(res$V2)~res$V1)
}
#' sbatch -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::runIDR()'
#' sbatch -p hipri test-job.sh
#' sbatch -p hipri test-job.sh
#' squeue -u steven.miamiuniv
runIDR <- function()
{
cmd="idr --samples /home/steven.miamiuniv/Softwares/idr-2.0.2/tests/data/peak1 /home/steven.miamiuniv/Softwares/idr-2.0.2/tests/data/peak2 --plot testIDR"
system(cmd)
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::peakCallAndAnnotationWithoutInput("/scratch/projects/bbc/aiminy_project/7_27_2017_bam/","/scratch/projects/bbc/aiminy_project/macs2_call","hs","macs2",0.0001)'
#
peakCallAndAnnotationWithoutInput <- function(input.file.dir,output.file.dir,genome = "hs",peakcaller = c("macs14", "macs2"), peakPvalue) {
genome <- match.arg(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
PATH1 = Sys.getenv("PATH")
macs2_Lib = file.path("/nethome/axy148/NGS_tools/MACS/bin/")
Sys.setenv(PATH = paste0(macs2_Lib, ":", PATH1))
cmd1 <- Sys.which("macs2")[[1]]
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-m 6,18 --bw=200 -p", peakPvalue, sep = " ")
})
re<-ParserReadFiles(input.file.dir,"bam")
file.name.2<-re$input
#output.dir.name=re$output
temp3=file.path(output.file.dir,"PeakCall")
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
re.out<-file.name.2
#cmd9="macs14 -t "
#cmd10="-f BAM -g hs -n "
#cmd11=" -m 6,18 --bw=200 -p 0.00001"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd12=paste(cmd9,x,cmd10,file.path(temp3,paste0(x_name,"_hs_1.00e-05_macs")),cmd11,sep=" ")
print(cmd12)
system(cmd12, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
#AnnotatePeak2(paste0(temp3,"/"),"*macs142_peaks.bed",7,paste0(output.dir.name,"PeakAnnotation_at_",temp2),genome="Hs")
#AnnotatePeak3(paste0(temp3,"/"),paste0(output.dir.name,"_PeakAnnotation"),
# genome="Hs")
#BamFileSortIndexVisualization2(re,genome)
}
#input <- "~/nodup.bam.txt"
#output <- "~/"
#
# R -e 'library(ChipSeq);library(ChipSeq);ChipSeq:::parseToSampleInfo("/projects/scratch/bbc/Project/Danny_chip3/Filtered_bam","*.bam$","~/Danny_chip3","sample_infor_Danny_chip3.txt","Danny_chip3")'
#
parseToSampleInfo <- function(input.dir,input.pattern,output.dir,output.file,label){
file.1 <- list.files(input.dir,pattern=input.pattern, all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
s <- file.1
ss <- basename(as.character(s))
sss <- tools::file_path_sans_ext(ss)
ssss <- tools::file_path_sans_ext(sss)
sm.info <- cbind.data.frame(sss,rep(label,length(sss)),ssss,rep("Link",length(sss)))
colnames(sm.info) <- c("ID","Type_Cell","Type_TF","Link")
if(!dir.exists(output.dir)){dir.create(output.dir,recursive = TRUE)}
write.table(sm.info,file=file.path(output.dir,output.file),row.names = FALSE,quote=FALSE,sep=",")
}
# ChipSeq:::getAb2inputPair("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt")
#
getAb2inputPair <- function(input.infor.file)
{
f <- read.table(input.infor.file,header = FALSE)
colnames(f)=c("Ab","Input")
print(f)
return(f)
}
# ChipSeq:::getBwUseBamCompare("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt","~")
#
getBwUseBamCompare <- function(input.infor.file,bl_file,output.dir)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
x.run <- apply(x.sample, 1, function(x,output.dir)
{
print(t(x))
x <- as.data.frame(t(x))
xx <- x$Ab
xx.input <- x$Input
#xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd1 <- paste("bamCompare --bamfile1",xx,"--bamfile2",xx.input,sep=" ")
cmd2 <- "--binSize 25"
#cmd3 <- "--ratio log2 -o"
cmd3 <- "--ratio subtract --normalizeTo1x 2451960000 --ignoreForNormalization chrX -o"
cmd3 <- "--ratio subtract --normalizeTo1x 2451960000 --ignoreForNormalization chrX --blackListFileName"
cmd3 <- paste(cmd3,bl_file,"-o",sep=" ")
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(xx.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
print(cmd6)
system(cmd6)
}, output.dir)
}
# R -e 'library(ChipSeq)::peakCall2("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt","hs","~/","macs2",0.00001)'
#
peakCall2 <- function(input.infor.file,genome = c("Hs","hs", "HS", "hS"),output.dir,peakcaller = c("macs14","macs2"), peakPvalue)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
peakcaller <- match.arg(peakcaller)
genome <- match.arg(genome)
genome <- tolower(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
cmd1 <- "macs2"
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-p", peakPvalue, sep = " ")
})
macs.run <- apply(x.sample,1,function(u, output.dir)
{
x <- as.data.frame(t(u))
xx <- x$Ab
xx.input <- x$Input
x.name <- paste(peakcaller,basename(as.character(xx)),basename(as.character(xx.input)),genome,peakPvalue,sep="_")
cmd12 = paste(cmd9, xx, "-c", xx.input, cmd10, file.path(output.dir, x.name), cmd11, sep = " ")
print(cmd12)
system(cmd12)
},output.dir)
#AnntationUsingChipSeeker(output.dir, "peaks.bed", temp3, DD = 5000)
# return(re)
}
useBamCoverage <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
#bamCoverage -b reads.bam -o coverage.bw
#bamCoverage --bam a.bam -o a.SeqDepthNorm.bw \
#--binSize 10
#--normalizeTo1x 2150570000
#--ignoreForNormalization chrX
#--extendReads
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX"
cmd3 <- "-o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("To1x_",basename(as.character(xx)),"_vs_","no_input",".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
getBwUseBamCoverage2 <- function(input.infor.file,bl_file,output.dir)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
x.run <- apply(x.sample, 1, function(x,output.dir)
{
print(t(x))
x <- as.data.frame(t(x))
xx <- x$Ab
xx.input <- x$Input
#xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd1 <- paste("bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX --blackListFileName"
cmd2 <- paste(cmd2,bl_file,sep=" ")
cmd3 <- "-o"
#cmd1 <- paste("bamCompare --bamfile1",xx,"--bamfile2",xx.input,sep=" ")
#cmd2 <- "--binSize 25"
#cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("Un_norma",basename(as.character(xx)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
print(cmd6)
system(cmd6)
}, output.dir)
}
getSummitSequence<-function(dir.name,input.file.pattern,genome,out.dir.name){
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.1 <- list.files(dir.name,pattern=input.file.pattern,all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
print(file.1)
re.out<-lapply(file.1,function(u){
#print(names(u))
uu=u
re=read.table(uu,header=TRUE)
print(head(re))
colnames(re)[c(7,9)]=c("-10*LOG10(pvalue)","FDR(%)")
temp<-re
temp$start<-temp$start-1
temp$end<-temp$end-1
temp$summit<-temp$abs_summit-1
temp
summitPeak<-temp$start+temp$summit
temp2<-temp
temp2$start<-summitPeak-49
temp2$end<-summitPeak+50
temp2$summit<-summitPeak
temp2$length<-temp2$end-temp2$start+1
temp2
rownames(temp2)<-paste0("MACS_peak_",rownames(temp2))
temp3<-toGRanges(temp2)
names(temp3)<-rownames(temp2)
if(genome == "mm10")
{
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Mmusculus.UCSC.mm10")
genome(temp3.trimmed)<-"mm10"
}
if(genome == "hg19"){
dd.GRCm39.mm10<-toGRanges(EnsDb.Hsapiens.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Hsapiens.UCSC.hg19")
genome(temp3.trimmed)<-"hg19"
}
seq.temp3<-getAllPeakSequence(temp3.trimmed,genome=genome.mm10)
write2FASTA(seq.temp3, file.path(out.dir.name,paste0(basename(u),".fa")))
#re2<-list(originalPeak=re,down1Peak=temp,aroundSummit100Peak=temp2,GR=temp3)
#re2
})
#sample.name<-sapply(strsplit(names(file.name.2),split="_peaks_"),"[[",1)
#names(re.out)<-sample.name
test
#return(re.out)
}
aiminy/ChipSeq documentation built on May 12, 2019, 3:37 a.m.
R Package Documentation
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#' AnnotatePeak2
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#' input.file.pattern="*macs142_peaks.bed"
#' output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#'
#' AnnotatePeak(input.file.dir,input.file.pattern,8,output.file.dir,genome="Mm")
#' AnnotatePeak(input.file.dir,input.file.pattern,7,output.file.dir,genome="Hs")
#'
#'
#'
AnnotatePeak2 <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",Sys.time())
#temp2=gsub(" ","-",temp1)
#temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
temp3=output.dir.name
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
#' AnnotatePeak3
#'
#' @param input.file.dir
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#' output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#'
#' AnnotatePeak3(input.file.dir,output.file.dir,genome="Mm")
#' AnnotatePeak3(input.file.dir,output.file.dir,genome="Hs")
#'
#'
#'
AnnotatePeak3<- function(input.file.dir,output.file.dir,genome) {
library(ChIPpeakAnno)
re<-ParserReadFiles(input.file.dir,"bed",output.file.dir)
#print(file.name.2)
file.name.2<-re$input
output.dir.name=output.file.dir
#dir.name=input.file.dir
#input.file.pattern=input.file.pattern
#dir.name=reformatPath(dir.name)
#output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",Sys.time())
#temp2=gsub(" ","-",temp1)
#temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
temp3=output.dir.name
dir.create(temp3)
#file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
#file.name.2<-as.list(file.name)
#names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
#print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
#' AnntationUsingChipSeeker
#'
#' Use ChipSeeker to annotate data
#'
#' @param dir.name: path for bed files
#' @param input.file.pattern : input file pattern
#' @param out.dir.name output file directory
#' @param txdb Annotation databased to be used("hg19","hg38")
#' @param DD distance definition around TSS
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed"
#' input.file.pattern="common_peaks.bed"
#' out.dir.name="/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnntationUsingChipSeeker <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=5000,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
#re<-ParserReadFiles(dir.name,input.file.pattern)
#re.bed<-re$input
#re.peaks.only.bed.2 <- re.bed
file.1 <- list.files(dir.name,pattern=input.file.pattern,all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
#re.bed<-re$input
re.peaks.only.bed.2 <- file.1
# if(length(dir(dir.name,pattern="peaks.bed"))!=0)
# {
# re.peaks.only.bed.2<-FL(re.bed,'peaks')
# cat("peaks\n")
# print(re.peaks.only.bed.2)
# }
#
# if(length(dir(dir.name,pattern="summits.bed"))!=0){
# re.summits.only.bed<-FL(re.bed,'summits')
# cat("summits\n")
# print(re.summits.only.bed)
# }
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,"Annotation",APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
res <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
#select.index <- which(peakAnno$distanceToTSS<20,000 && peakAnno$distanceToTSS >= -20,000)
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
x_name=names(re.peaks.only.bed.2)[u]
cat(x_name)
png(file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_pie.png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
#print(paste0(peaks[,c(2,3)]))
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_only_mapped_peaks.xls")),
row.names = FALSE,quote=FALSE,sep="\t")
unmapped.peaks<-peaks[-which(paste0(peaks[,2],"_",peaks[,3]) %in% paste0(peaks.anno[,2],"_",peaks.anno[,3])),]
cat(dim(peaks)[1]," ",dim(peaks.anno)[1]," ",dim(unmapped.peaks)[1],"\n")
if(dim(unmapped.peaks)[1]!=0){
colnames(unmapped.peaks)=colnames(peaks.anno)[1:6]
unmapped.peaks.3<-smartbind(peaks.anno,unmapped.peaks)
unmapped.peaks.4<-unmapped.peaks.3[order(unmapped.peaks.3[,1],unmapped.peaks.3[,2]),]
write.table(unmapped.peaks.4,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_all_peaks.xls")),row.names = FALSE,quote=FALSE,sep="\t")
}
re <- list(peaksMappedOnly=peaks.anno,peaksAll=unmapped.peaks.4)
re
},re.peaks.only.bed.2,d)
res
}
#' BamFileSortIndexVisualization
#'
#' @param input.file.dir
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' output.file.dir="/scratch/projects/bbc/aiminy_project/"
#' genome="Hs"
#' d=4000
#' BamFileSortIndexVisualization(input.file.dir,output.file.dir,4000,genome)
#'
#'bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::BamFileSortIndexVisualization("/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA","/scratch/projects/bbc/aiminy_project/7_27_2017",400000,"Hs")'
#'
#'
BamFileSortIndexVisualization <- function(input.file.dir,input.pattern,output.file.dir,d,genome,bam.sort=FALSE) {
#library(ChIPpeakAnno)
#re<-ParserReadFiles(input.file.dir,"bam")
file.1 <- list.files(input.file.dir,pattern=input.pattern, all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
file.name.2<-file.1
#output.dir.name=re$output
#temp3=paste0(output.dir.name,"_visualization")
temp3=output.file.dir
if (!dir.exists(temp3))
{
dir.create(temp3, recursive = TRUE)
}
#dir.create(temp3)
re.out<-file.name.2
if(bam.sort==FALSE){
cmd1="samtools sort"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd2=paste0(cmd1," ",x," ","-o"," ",file.path(temp3,paste0(x_name,"_sorted.bam")))
print(cmd2)
system(cmd2)
},re.out,temp3)
cmd3="samtools index"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd4=paste0(cmd3," ",file.path(temp3,paste0(x_name,"_sorted.bam")))
#print(cmd2)
system(cmd4)
},re.out,temp3)
}
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
#cmd7="-L 4000"
cmd7=paste("-L",d,sep=" ")
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=tools::file_path_sans_ext(basename(x))
cmd8=paste(cmd5,file.path(temp3,paste0(x_name,"_sorted.bam")),cmd6,file.path(temp3,x_name),cmd7,sep=" ")
print(cmd8)
system(cmd8, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
}
convertBam2StrandBw2 <- function(input.bam.file.dir, output.bw.file.dir, BigMem = FALSE,
cores = 15, Memory = 25000, Wall.time = "72:00", span.ptile = 8)
{
re <- parserreadfiles(input.bam.file.dir, "bam")
res <- re$input
m.id <- grep("login", system("hostname", intern = TRUE))
if (!dir.exists(output.bw.file.dir))
{
dir.create(output.bw.file.dir, recursive = TRUE)
}
# job.name=paste0('bamSort[',length(res),']')
cmd.l <- lapply(1:length(res), function(u, m.id, Wall.time, cores, Memory,
span.ptile, res, output.bw.file.dir)
{
# path_name = dirname(res[[u]]) path_name2 <- basename(path_name)
file_name = file_path_sans_ext(basename(res[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
if (BigMem == TRUE)
{
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
} else
{
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' aimin.yan@med.miami.edu",
sep = " ")
}
job.name = paste0("bam2wig.", u)
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.",
job.name, ".log "), paste0("-e %J.", job.name, ".err -W"))
# job.name=paste0('bamSort[',length(res),']') cmd1 = paste0('bsub -w
# \'done(\'bamSort[*]\')\'', 'bsub -P bbc -J \'',job.name,paste0('\'
# -o %J.log '),paste0('-e %J.err -W')) job.name=paste0('Bdg[',u,']') cmd1 =
# paste0('bsub -w \'done(\'bamIndex[*]\') && done(\'Chrosome\')\'',
# 'bsub -P bbc -J \'',job.name,paste0('\' -o %J.',job.name,'.log
# '),paste0('-e %J.',job.name,'.err -W'))
if (u <= 6)
{
cmd2 = paste("bam2wig.pl -pe --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
} else
{
cmd2 = paste("bam2wig.pl --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
}
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
if (u <= 6)
{
cmd3 = paste("bam2wig.pl -pe --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
} else
{
cmd3 = paste("bam2wig.pl --pos span --strand --bw --bwapp $HOME/kentUtils/bin/wigToBigWig --out",
file.path(output.bw.file.dir, paste0(file_name, "_2.bw")),
"--in", res[[u]], sep = " ")
}
}
cmd <- cmd3
system(cmd)
cmd
}, m.id, Wall.time, cores, Memory, span.ptile, res, output.bw.file.dir)
}
#'R -e 'library(ChipSeq);ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot")'
#'R -e 'library(ChipSeq); x <- ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot",cores = 8, Memory = 16000,span.ptile = 4,wait = FALSE)'
#'R -e 'library(ChipSeq); x <- ChipSeq:::plotBam(input.file.dir="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA",file.type="*marked.bam",output.file.dir="/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot",job.option = "parallel", cores = 32, Memory = 16000,span.ptile = 16,wait = TRUE)'
plotBam <- function(input.file.dir,file.type,output.file.dir,job.option = "general",cores = 15, Memory = 25000, Wall.time = "72:00", span.ptile = 8,wait=TRUE) {
#library(ChIPpeakAnno)
#cat(job.option,"\n")
#job.option <- as.character(job.option)
#cat(job.option,"\n")
re<-ParserReadFiles(input.file.dir,file.type)
file.name.2<-re$input
#output.dir.name=re$output
re.out<-file.name.2
m.id <- grep("login", system("hostname", intern = TRUE))
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp3= output.file.dir
#cmd1="samtools sort"
cmd.1 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamSort.", u)
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.",
job.name, ".log "), paste0("-e %J.", job.name, ".err -W"))
cmd2=paste("samtools sort",re.out[[u]],file.path(temp3,paste0(file_name, "_sorted")),sep=" ")
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
cmd3 = paste("samtools sort",re.out[[u]],file.path(temp3,paste0(file_name, "_sorted")),sep=" ")
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
cmd.2 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamIndex.", u)
wait.job.name = paste0("bamSort.", u)
if(wait == TRUE){
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
}else{
cmd1 = paste0("bsub -P bbc -J \"",job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
}
cmd2=paste("samtools index",file.path(temp3,paste0(file_name, "_sorted.bam")),sep=" ")
cmd3 = paste(cmd1, cmd0, cmd2, sep = " ")
} else
{
cmd3 = paste("samtools index",file.path(temp3,paste0(file_name, "_sorted.bam")),sep=" ")
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
cmd.3 <- lapply(1:length(re.out),function(u,m.id,job.option,Wall.time, cores, Memory,
span.ptile,re.out,temp3){
file_name = file_path_sans_ext(basename(re.out[[u]]))
# file_name <- paste0(path_name2,'-',file_name)
#u <- 3
if (m.id == 1)
{
#job.option <- match.arg(job.option)
switch (job.option,
parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
},
general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}
)
job.name = paste0("bamPlot.", u)
wait.job.name = paste0("bamIndex.", u)
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
cmd8=paste(cmd5,file.path(temp3,paste0(file_name,"_sorted.bam")),cmd6,file.path(temp3,paste0(file_name,"_sorted")),cmd7,sep=" ")
cmd3 = paste(cmd1, cmd0, cmd8, sep = " ")
} else
{
"ngs.plot.r -G hg19 -R tss -C config.txt -O 2017-03-02-01_S5_R1.tss -T 2017-03-02-01_S5_R1 -L 4000 -RR 1"
"ngs.plot.r -G hg19 -R tss -C config.txt -O 2017-03-02-01_S5_R1.tss -T 2017-03-02-01_S5_R1 -L 4000 -RR 1 -GO max -SC global"
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
cmd8=paste(cmd5,file.path(temp3,paste0(file_name,"_sorted.bam")),cmd6,file.path(temp3,paste0(file_name,"_sorted")),cmd7,sep=" ")
cmd3 = cmd8
}
cmd <- cmd3
system(cmd)
cmd
},m.id,job.option,Wall.time,cores,Memory,span.ptile,re.out,temp3)
#re <- list(cmd.1 = cmd.1,cmd.2=cmd.2,cmd.3=cmd.3)
# return(re)
}
#' DrawVenn
#'
#' @param devtools
#' @param install_github
#' @param Vennerable
#' @param venneuler
#' @param VennDiagram
#'
#' @return
#' @export
#'
#' @examples
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#' DrawVenn(out.dir.name)
#'
DrawVenn <- function(out.dir.name) {
devtools::install_github("js229/Vennerable");
library(Vennerable)
BB=c(1:6153,6154:25736)
CC=c(6154:25736,25737:29579)
AA=c(6026:6153,6154:8462,25737:25776,29650:29719)
Vstem <- Venn(list(ASXL1=AA,SMC1A=BB,RAD21=CC))
SetLabels <- VennGetSetLabels(Vstem)
Cstem3 <- compute.Venn(Vstem,doWeights=TRUE)
SetLabels <- VennGetSetLabels(Cstem3)
FaceLabels <- VennGetFaceLabels(Cstem3)
FaceLabels[FaceLabels$FaceName=="010","x"] <- 10
FaceLabels[FaceLabels$FaceName=="010","y"] <- 3
SetLabels[SetLabels$Label=="SMC1A","hjust"] <- "right"
SetLabels[SetLabels$Label=="SMC1A","x"] <- 110
SetLabels[SetLabels$Label=="SMC1A","y"] <- 58.78653
SetLabels[SetLabels$Label=="RAD21","x"] <- -80
SetLabels[SetLabels$Label=="RAD21","y"] <- 58.78653
SetLabels[SetLabels$Label=="ASXL1","y"] <-80
Cstem3 <- VennSetSetLabels(Cstem3,SetLabels)
Cstem3<-VennSetFaceLabels(Cstem3,FaceLabels)
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="010"),]$x<-90
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="010"),]$y<--35
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="001"),]$x<--90
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="001"),]$y<--35
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$x<--18
#Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$y<--
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="101"),]$vjust<-"center"
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="110"),]$x<-19
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="110"),]$y<-49.86767
c.3.set=sum(Cstem3@IndicatorWeight[,4])
Cstem3@IndicatorWeight[which(row.names(Cstem3@IndicatorWeight)=="000"),4]=35000-c.3.set
Cstem3@FaceLabels[which(Cstem3@FaceLabels$FaceName=="DarkMatter"),]$y<--130
temp<-ol$venn_cnt
index.A=grep(colnames(Cstem3@IndicatorWeight)[1],colnames(temp))
index.B=grep("SMC1",colnames(temp))
index.C=grep("Rad21",colnames(temp))
temp2<-as.data.frame(temp[,c(index.A,index.B,index.C,4)])
colnames(temp2)[1:3]=colnames(Cstem3@IndicatorWeight)[1:3]
row.names(temp2)<-with(temp2,paste0(temp2$ASXL1,temp2$SMC1A,temp2$RAD21))
row.names(temp2)<-row.names(Cstem3@IndicatorWeight)
temp3<-merge(Cstem3@IndicatorWeight,temp2,by=0,sort=FALSE)
Cstem3@IndicatorWeight[,4]<-temp3$Counts
c.3.set=sum(Cstem3@IndicatorWeight[,4])
Cstem3@IndicatorWeight[which(row.names(Cstem3@IndicatorWeight)=="000"),4]=35000-c.3.set
grid.newpage()
pdf(paste0(out.dir.name,"venn.pdf"))
plot(Cstem3)
dev.off()
}
#' GetResultsFromDiffBind
#'
#' Use DiffBind to process peak profiles
#'
#' @param mcf7
#' @param Mergereplicates
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#' output.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff_7_21_2017_2"
#' mcf7=resmcf
#' re<-GetResultsFromDiffBind(mcf7,"yes",output.file.dir)
#' re<-GetResultsFromDiffBind(mcf2,"yes",output.file.dir)
#'
GetResultsFromDiffBind<-function(mcf7,Mergereplicates=c("yes","no"),output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp<-mcf7
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
if(Mergereplicates=="yes"){
temp2<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp2)
##need to set the flexiable number identified from data sets
t <- temp2
temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
}else{
temp22<-temp
}
print(temp22)
Tissue1<-temp22$class[2,]
Tissue2<-unique(temp22$class[2,])
TF<-unique(temp22$class[3,])
TF.n<-length(TF)
temp3=file.path(output.file.dir,"Venn")
if(!dir.exists(temp3))
{
dir.create(temp3)
}
for(i in 1:length(Tissue2)){
po<-which(Tissue1 %in% Tissue2[i])
print(po)
if(length(po)==2)
{
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po,main=Tissue2[i])
dev.off()
}else if(length(po)==4){
po1<-po[c(1,3)]
po2<-po[c(2,4)]
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
dev.off()
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
dev.off()
}else
{
cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
}
}
p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
po<-which(Tissue1 %in% Tissue2[u])
if(length(po)==2)
{
common.peaks<-dba.overlap(temp22,mask=po)
y<-common.peaks$inAll
}else if(length(po)==4){
po1<-po[c(1,3)]
po2<-po[c(2,4)]
common.peaks.1<-dba.overlap(temp22,mask=po1)
y1<-common.peaks.1$inAll
common.peaks.2<-dba.overlap(temp22,mask=po2)
y2<-common.peaks.2$inAll
y<-list(y1=y1,y2=y2)
names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
}else{
y<-NULL}
y
},Tissue1,Tissue2,temp22)
names(p.common)<-Tissue2
p.common<-unlist(p.common,recursive = F)
p.common<-p.common[lapply(p.common,length) > 0]
if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
{
dir.create(file.path(output.file.dir,"common_peaks_bed"))
dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
}
#output common peaks to bed files
lapply(1:length(p.common),function(u,p.common,output.file.dir){
x=p.common[[u]]
x_name=names(p.common)[u]
df <- data.frame(seqnames=seqnames(x),
#starts=start(x)-1,
starts=start(x),
ends=end(x),
names=c(rep(".", length(x))),
scores=elementMetadata(x)[,1],
strands=strand(x))
#assign strand
df.str <- data.frame(seqnames=seqnames(x),
#starts=start(x)-1,
starts=start(x),
ends=end(x),
names=c(rep(".", length(x))),
scores=elementMetadata(x)[,1],
strands=c(rep(".", length(x))))
df.str.1<-df.str[-grep("random",df.str$seqnames),]
df.str.2<-df.str.1
df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
},p.common,output.file.dir)
AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
return(p.common)
}
#' GetSampleInfo
#'
#' GetSampleInfo
#'
#' @param input.sample.file
#' @param input.bam.file
#'
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/sorted_bam_files.txt'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny/sorted_bam_files_2.txt'
#'
#' re <- GetSampleInfo(input.sample.file,input.bam.file)
#'
#'
GetSampleInfo <- function(input.sample.file, input.bam.file)
{
re <- read.csv(input.sample.file)
re.c <- colnames(re)
bam.file <- read.table(input.bam.file, header = FALSE)
yy <- apply(bam.file, 1, function(u)
{
y <- basename(u)
pos <- regexpr("\\.", y)
pos <- pos - 1
y <- substr(y, 1, pos)
y
})
bam.file.sample.name <- cbind(bam.file, yy)
colnames(bam.file.sample.name) <- c("file.name", "ID")
Ab.type <- unique(as.character(re[, colnames(re) == re.c[3]]))
Cell.type <- unique(as.character(re[, colnames(re) == re.c[2]]))
re1 <- cbind(re[, 1:3], paste0(re[, 2], "_", re[, 3]))
re11 <- merge(re1, bam.file.sample.name, by = "ID")
colnames(re11)[4] = "Cell_TF"
chipseqCell <- function(n, es)
{
value <- list(name = n, es = es)
attr(value, "class") <- "ChipSeqCell"
value
}
cellType <- unique(as.character(re1[re1$Type_TF == "Input", ]$Type_Cell))
yy <- lapply(cellType, function(u, re11)
{
n <- nchar(u)
index1 <- which(substr(re11$Type_Cell, 1, n) == u)
z <- re11[index1, ]
index2 <- which(nchar(as.character(z$Type_Cell)) <= n + 2)
z2 <- z[index2, ]
cc <- chipseqCell(u, z2)
cc
}, re11)
colnames(re1)[4] = "Cell_TF"
Cell.Ab.type <- unique(as.character(re1[, 4]))
re2 <- lapply(1:length(Cell.Ab.type), function(u, Cell.Ab.type, re1)
{
x = Cell.Ab.type[u]
z = re1
ZZ <- as.character(z[which(z[, 4] == x), 1])
ZZ
}, Cell.Ab.type, re1)
names(re2) <- Cell.Ab.type
re21 <- re2[lapply(re2, length) == 1]
re22 <- re2[lapply(re2, length) == 2]
re3 <- list(re1 = re1, re2 = re2, re21 = re21, re22 = re22, re11 = re11,
y = yy)
return(re3)
}
#' peakcallwithinput
#'
#' @export
#' @example
#'
#' genome='Hs'
#' re <- peakcallwithinput(input.sample.file,input.bam.file,genome,output.dir,peakcaller)
#'
#'
peakcallwithinput <- function(input.sample.file, input.bam.file, genome = c("Hs",
"hs", "HS", "hS"), output.dir, peakcaller = c("macs14", "macs2"), peakPvalue)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
output.dir.name = dirname(input.sample.file)
temp3 = file.path(output.dir.name, output.dir)
if (!dir.exists(temp3))
{
dir.create(temp3)
}
peakcaller <- match.arg(peakcaller)
genome <- match.arg(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
PATH1 = Sys.getenv("PATH")
macs2_Lib = file.path("/nethome/axy148/NGS_tools/MACS/bin/")
Sys.setenv(PATH = paste0(macs2_Lib, ":", PATH1))
cmd1 <- Sys.which("macs2")[[1]]
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-p", peakPvalue, sep = " ")
})
cellInfo.run <- lapply(1:length(cellInfo), function(u, cellInfo, temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
x.run <- apply(x.sample, 1, function(x)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd12 = paste(cmd9, xx, "-c", x.input, cmd10, file.path(temp3, paste0(xx.name,
"_hs_1.00e-05_", peakcaller)), cmd11, sep = " ")
cmd12
})
x.run
}, cellInfo, temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
re <- list(cellInforun = cellInfo.run, zzz = zzz)
AnntationUsingChipSeeker(temp3, "peaks.bed", temp3, DD = 5000)
return(re)
}
select.sample <- c("MDA MB 231-DD-1_cJun", "MDA MB 231-1_cJun", "1833-1_cJun")
output.config.dir <- "~/"
configAndMultiplot <- function(res, select.sample, output.config.dir)
{
if (!dir.exists(output.config.dir))
{
dir.create(output.config.dir)
}
x <- res$re11
xx <- x[which(x$Cell_TF %in% select.sample), ]
xxx <- cbind.data.frame(xx$file.name, rep("-1", dim(xx)[1]), gsub(" ", "-",
xx$Cell_TF))
config.sample.name <- paste(gsub(" ", "-", select.sample), collapse = "-and-")
config.file.name <- file.path(output.config.dir, paste0(config.sample.name,
"-config.txt"))
write.table(xxx, file = config.file.name, col.names = FALSE, row.names = FALSE,
quote = FALSE, sep = "\t")
cmd0 <- "ngs.plot.r -G hg19 -R tss -C"
cmd1 <- "-O"
cmd2 <- "-L 4000 -RR 1 -CD 1 -CO \"blue\""
output.results <- file.path(output.config.dir, paste0(config.sample.name,
"results"))
cmd <- paste(cmd0, config.file.name, cmd1, output.results, cmd2, sep = " ")
system(cmd)
return(config.file.name)
}
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny/sorted_bam_files_2.txt'
#'
#' re <- ChipSeq:::matchBamInputGene(input.sample.file,input.bam.file,'$HOME/all_common_gene_unique.txt','$HOME/NgsConfigFile')
#'
#' R -e 'libraray(ChipSeq);re <- ChipSeq:::matchBamInputGene('/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv','/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt','$HOME/all_common_gene_unique.txt','$HOME/NgsConfigFile',,job.option = 'parallel')'
#'
#' bsub -P bbc -J 'bamPlot' -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::matchBamInputGene('/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv','/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt','~/all_common_gene_unique.txt','NgsConfigFile')'
#'
#' bsub -P bbc -J "bamPlot" -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::matchBamInputGene("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt","~/all_common_gene_unique.txt","NgsConfigFile3",add.input= "yes")'
matchBamInputGene <- function(input.sample.file, input.bam.file, input.gene.list,
output.dir, ngs.para = c("hg19", 4000, 1, 1, "total"), add.input = NULL)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u, add.input, cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
x.run <- apply(x.sample, 1, function(x, add.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
if (!is.null(add.input))
{
cmd12 = paste(paste0(xx, ":", x.input), input.gene.list, paste0(gsub(" ",
"-", Cell_TF), "_cJunAndp27"), sep = "\t")
} else
{
cmd12 = paste(xx, input.gene.list, paste0(gsub(" ",
"-", Cell_TF), "_cJunAndp27"), sep = "\t")
}
cat(cmd12, file = file.path(temp3, paste0(xx.name, "_config_cJunAndp27.txt")),
sep = "\t")
}, add.input, temp3)
x.run
}, add.input, cellInfo, temp3)
# dir.name=temp3 dir.name=reformatPath(dir.name)
file.name = file.path(temp3, dir(temp3, recursive = TRUE))
file.name.2 <- as.list(file.name)
# names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
zzz <- unlist(file.name.2)
lapply(1:length(zzz), function(u, zzz)
{
dir.name = dirname(zzz[u][[1]])
file_name = file_path_sans_ext(basename(zzz[u][[1]]))
cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
sep = " ")
# system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
cmd2 = cmd
cat(cmd2, "\n")
cat("\n")
system(cmd2)
}, zzz)
re <- list(cellInforun = cellInfo.run, zzz = zzz)
# AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
return(re)
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "bamCompare" -o %J.bamCompare.log -e %J.Compare.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCompare("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/scratch/projects/bbc/aiminy_project/DannyNewData2/sorted_bam_files_2.txt","~/BamCompare")'
useBamCompare <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1",xx,"--bamfile2",x.input,sep=" ")
cmd2 <- "--binSize 25"
cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(x.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
sysyem("module unload python/2.7.3")
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
#' x <- ChipSeq:::usePegasus('parallel', Wall.time = '72:00',cores = 32,Memory = 16000,span.ptile = 16,'bamPlot')
usePegasus <- function(job.option = c("general", "parallel", "bigmem"), Wall.time,
cores, Memory, span.ptile, job.name, wait.job.name = NULL)
{
job.option <- match.arg(job.option)
switch(job.option, parallel = {
cmd0 = paste(Wall.time, "-n", cores, "-q parallel -R 'rusage[mem=",
Memory, "] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu",
sep = " ")
}, bigmem = {
cmd0 = paste(Wall.time, "-n", cores, "-q bigmem -R 'rusage[mem=", Memory,
"] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu", sep = " ")
}, general = {
cmd0 = paste(Wall.time, "-n", cores, "-q general -R 'rusage[mem=", Memory,
"] span[ptile=", span.ptile, "]' -u aimin.yan@med.miami.edu", sep = " ")
})
if (!is.null(wait.job.name))
{
cmd1 = paste0("bsub -w \"done(\"", wait.job.name, "\")\"", " -P bbc -J \"",
job.name, paste0("\" -o %J.", job.name, ".log "), paste0("-e %J.",
job.name, ".err -W"))
} else
{
cmd1 = paste0("bsub -P bbc -J \"", job.name, paste0("\" -o %J.", job.name,
".log "), paste0("-e %J.", job.name, ".err -W"))
}
cmd = paste(cmd1, cmd0, sep = " ")
return(cmd)
}
#' IndexBamFile
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' input.file.pattern="*.bam"
#' index.file=9
#' output.file.dir="/scratch/projects/bbc/aiminy_project/
#' genome="Hs"
#'
#' IndexBamFile(input.file.dir,input.file.pattern,index.file,output.file.dir,genome)
#'
IndexBamFile <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
#library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
temp=Sys.time()
temp1=gsub(":","-",Sys.time())
temp2=gsub(" ","-",temp1)
temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
#print(file.name.2)
re.out<-file.name.2
cmd1="samtools index "
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd2=paste0(cmd1," ",x," ",paste0(temp3,"/",x_name,"_sorted"))
#print(cmd2)
system(cmd2)
},re.out,temp3)
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
#file.name.4 <-file.name.3[-1]
#re.out<-lapply(file.name.4,function(u){
# re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
# re
#})
#head(re.out[[1]])
# re.out.L<-lapply(re.out,function(u){
# re=length(u)
# #colnames(re)=c("Count","GeneName")
# re
# })
#
#
# # cmd="samtools sort"
# # input.file="/scratch/projects/bbc/Project/Danny_chip2/Alignment/BWA/""2016-10-14-Hyunho-Yoon-16-1833-PF1502-1-p27_S4_R1.bam"
# # output.file="/scratch/projects/bbc/aiminy_project/Bam_marked_sorted/""2016-10-14-Hyunho-Yoon-16-1833-PF1502-1-p27_S4_R1.bam_sorted"
#
# if(genome=="Mm"){
#
# annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
#
# ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
#
# overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
#
# binOverFeature(overlaps, annotationData=annoData,
# radius=5000, nbins=20, FUN=length, errFun=0,
# ylab="count",
# main="Distribution of aggregated peak numbers around TSS")
#
# overlaps.trimmed<-trim(overlaps, use.names=TRUE)
#
# library(EnsDb.Mmusculus.v79)
# dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
# #seqinfo(dd.GRCm39.mm10)
# #seqlevels(dd.GRCm39.mm10)
#
# seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#
# seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
# #GRCm38/mm10
# #dd<-toGRanges(EnsDb.Mmusculus.v79)
# #seqinfo(dd)
# #library(ensembldb)
# #library(GenomeInfoDb)
# seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
# overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
# write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
# }else if(genome=="Hs"){
#
# library(EnsDb.Hsapiens.v75)
# #annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
#
# dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
#
# print(seqinfo(dd.hs))
# print(seqlevels(dd.hs))
#
# #print(seqlevels(dd.hs)[,1])
#
# #print(seqlevels(re.out[[1]])[,1])
#
# # seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# # "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# # "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#
# #temp4=
#
# re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
#
# x=re.out[[u]]
# x_name=names(re.out)[u]
#
# seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
# #print(seqinfo(re.out.trimmed))
# #print(seqlevels(re.out.trimmed))
# seqinfo(x)<-seqinfo(dd.hs)
# #GRCm38/mm10
# #dd<-toGRanges(EnsDb.Mmusculus.v79)
# #seqinfo(dd)
# #library(ensembldb)
# #library(GenomeInfoDb)
# seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
# re.out.trimmed<-trim(x, use.names=TRUE)
# overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
#
# write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
# },re.out,dd.hs)
#
# }
}
#' InstallRequiredPackage
#'
#' @return
#' @export
#'
#' @examples
#' InstallRequiredPackage()
#'
InstallRequiredPackage <- function() {
#install from bioc
source("http://bioconductor.org/biocLite.R")
biocLite("GenomicRanges")
biocLite("ChIPseeker")
biocLite("regioneR")
biocLite("DiffBind")
biocLite("BSgenome.Hsapiens.UCSC.hg19")
biocLite("TxDb.Mmusculus.UCSC.mm9.knownGene")
biocLite("EnsDb.Hsapiens.v75")
devtools::install_github("sheffien/simpleCache")
biocLite("LOLA")
biocLite("ChIPpeakAnno")
biocLite("BSgenome.Mmusculus.UCSC.mm9")
biocLite("motifStack")
biocLite("BSgenome.Ecoli.NCBI.20080805")
biocLite("EnsDb.Mmusculus.v75")
biocLite("EnsDb.Mmusculus.v79")
biocLite("BSgenome.Mmusculus.UCSC.mm10")
biocLite("org.Mm.eg.db")
#install from git
devtools::install_github("nsheff/LOLA")
#loading
library(BiocInstaller)
library(BiocInstaller)
library(ChIPseeker)
library(DiffBind)
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
library(TxDb.Mmusculus.UCSC.mm9.knownGene)
library(LOLA)
library("simpleCache")
library(ChIPpeakAnno)
library(rtracklayer)
library(org.Mm.eg.db)
library(motifStack)
library(BSgenome.Mmusculus.UCSC.mm10)
library(BSgenome.Mmusculus.UCSC.mm9)
library(EnsDb.Mmusculus.v75)
library(EnsDb.Mmusculus.v79)
library(ggplot2)
}
#' Title
#'
#' @return
#' @export
#'
#' @examples
#' LoadRequiredPackage()
#'
LoadRequiredPackage <- function() {
library(LOLA)
library(ChIPseeker)
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
library(clusterProfiler)
library(regioneR)
library(BSgenome.Hsapiens.UCSC.hg19)
}
#' ParserBamFile4NgsPlot
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#'
#' sh ~/Code/BashRunMACS1-4-2_4_Danny_chip_seq3.sh /scratch/projects/bbc/aiminy_project/Bam_sorted/ 7 ".sorted.bam$" /scratch/projects/bbc/aiminy_project/Bam_marked_sorted/
#'
#' dir.name="/scratch/projects/bbc/aiminy_project/Bam_sorted/"
#' index=7
#' input.file.pattern=".sorted.bam$"
#' out.dir.name="/scratch/projects/bbc/aiminy_project/Bam_marked_sorted/"
#' outfile="test.txt"
#'
#' re.from.bed.peng.4.venn<-ParserBamFile4NgsPlot(dir.name,index,input.file.pattern,out.dir.name,outfile)
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBamFile4NgsPlot<-function(dir.name,index,input.file.pattern,out.dir.name,outfile){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
#file.name="/scratch/projects/bbc/Project/Danny_chip/Alignment/BWA/2016-10-14-Hyunho-Yoon-02-MDA-MB-231-2-cJun_S5_R1.marked.bam"
#strsplit(file.name,split="\\/")
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index)
print(file.name.2)
ab<-names(file.name.2)
#ab<-"2016-10-14-Hyunho-Yoon-13-MDA-MB-231-DD-2-p27_S15_R1.marked.bam"
ab2<-sapply(strsplit(ab,split="\\."),"[[",1)
print(ab2)
#ab3<-sapply(strsplit(ab2,split="\\-"),"[[",length(strsplit(ab2,split="\\-")[[1]]))
re<-cbind(paste0(dir.name,names(file.name.2)),rep(-1,length(names(file.name.2))),ab2)
write.table(re, file =paste0(out.dir.name,outfile), append = FALSE, quote = F, sep = " ",eol = "\n", na = "NA", dec = ".", row.names = F, col.names = F)
cmd1="ngs.plot.r -G hg19 -R tss -C"
cmd2="-O"
cmd3="-L 4000"
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
cmd4=paste(cmd1,u,cmd2,u,cmd3,sep=" ")
print(cmd4)
system(cmd4, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
})
#sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
#names(re.out)=sample.name
# A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
# B=re.out[[2]] #10_WT_BM_ASXL1
# C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
# D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
#
# A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
# B.name=sample.name[2] #10_WT_BM_ASXL1
# C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
# D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#
# #A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
# #B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
# #class(A)
# #class(B)
#
#
# # TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
# # NumOverLap<-numOverlaps(A, B, count.once=TRUE)
# # pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
# # png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
# # plot(pt)
# # dev.off()
# #
# # lz <- localZScore(pt=pt, A=A, B=B)
# # png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
# # plot(lz)
# # dev.off()
# # }
# #
# # TestOverlap(A,B,A.name,B.name,out.dir.name,100)
# # TestOverlap(A,C,A.name,C.name,out.dir.name,100)
# # TestOverlap(A,D,A.name,D.name,out.dir.name,100)
# #
# # TestOverlap(B,C,B.name,C.name,out.dir.name,100)
# # TestOverlap(B,D,B.name,D.name,out.dir.name,100)
# # TestOverlap(C,D,C.name,D.name,out.dir.name,100)
# #
#
# # temp.name<-strsplit(names(file.name.2),split="\\.")
#
# #a = readBed(system.file("extdata", "examples/combined_regions.bed",
# # package="LOLA"))
#
#
# venn.plot <- venn.diagram(
# x = re.out[c(1,2)],
# filename = paste0(out.dir.name,names(re.out)[1],names(re.out)[2],"_overlap_venn.tiff"),
# height = 3000,
# width = 3500,
# resolution = 1000,
# col = "black",
# lty = "dotted",
# lwd = 1,
# fill = c("red","blue"),
# alpha = 0.50,
# label.col = c(rep("white",3)),
# cex = 0.5,
# fontfamily = "serif",
# fontface = "bold",
# cat.col = c("red","blue"),
# cat.cex = 0.5,
# cat.pos = 0.5,
# cat.dist = 0.05,
# cat.fontfamily = "serif"
# )
#
# venn.plot <- venn.diagram(
# x = re.out[c(1,3,5)],
# filename =paste0(out.dir.name,names(re.out)[1],names(re.out)[3],names(re.out)[5],"_overlap_venn.tiff"),
# col = "black",
# lty = "dotted",
# lwd = 2,
# fill = c("red", "orange", "blue"),
# alpha = 0.50,
# label.col = c(rep("white",7)),
# cex = 1,
# fontfamily = "serif",
# fontface = "bold",
# cat.col = c("red", "orange", "blue"),
# cat.cex = 0.8,
# cat.fontfamily = "serif"
# )
#return(re.out)
}
#' Title
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeakCall/"
#' input.file.pattern="*bam_mm_summits_from_PeakCall4Yang.bed"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#' output.test.file= "SMC1A_RAD21_overlap_test"
#'
#'
#' dir.name.2="/media/H_driver/2016/Yang/BedFromPeakCall_mm_100e_07/"
#' input.file.pattern.2="*_bam_mm_1_from_PeakCall4Yang.bed"
#'
#' out.dir.name.2="/media/H_driver/2016/Yang/Results/"
#' output.test.file.2= "SMC1A_RAD21_mm_100e-07_peaks_overlap_test"
#'
#' ParserBedFile(dir.name,input.file.pattern,out.dir.name)
#'
#' re.mm.100e.07<-ParserBedFile(dir.name.2,input.file.pattern.2,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_2.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_2.Rhistory"))
#'
ParserBedFile<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.3,function(u){
re=readBed(u)
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(sapply(strsplit(names(re.out),split="_i"),"[[",1),split="2470IUPUI_"),"[[",2)
A=re.out[[2]] #SMC1
B=re.out[[4]] #Rad21
C=re.out[[1]] #ASXL1
A.name=sample.name[2] #SMC1 name
B.name=sample.name[4] #Rad21 name
C.name=sample.name[1] #ASXL1 name
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
TestOverlap <- function(A, B, A.name,B.name,out.dir.name) {
NumOverLap<-numOverlaps(A, B, count.once=TRUE)
pt <- overlapPermTest(A=A, B=B, ntimes=50)
png(filename=paste0(out.dir.name,A.name,B.name,"_overlap_test.png"))
plot(pt)
dev.off()
lz <- localZScore(pt=pt, A=A, B=B)
png(filename=paste0(out.dir.name,A.name,B.name,"_localZScore.png"))
plot(lz)
dev.off()
}
TestOverlap(A,B,A.name,B.name,out.dir.name)
TestOverlap(A,C,A.name,C.name,out.dir.name)
TestOverlap(B,C,B.name,C.name,out.dir.name)
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
return(re.out)
}
#' ParserBedFile4Peng
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeng/"
#' input.file.pattern="*.bed"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng<-ParserBedFile4Peng(dir.name,input.file.pattern,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4Peng<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
re=readBed(u)
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
B=re.out[[2]] #10_WT_BM_ASXL1
C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
B.name=sample.name[2] #10_WT_BM_ASXL1
C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
NumOverLap<-numOverlaps(A, B, count.once=TRUE)
pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
plot(pt)
dev.off()
lz <- localZScore(pt=pt, A=A, B=B)
png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
plot(lz)
dev.off()
}
TestOverlap(A,B,A.name,B.name,out.dir.name,100)
TestOverlap(A,C,A.name,C.name,out.dir.name,100)
TestOverlap(A,D,A.name,D.name,out.dir.name,100)
TestOverlap(B,C,B.name,C.name,out.dir.name,100)
TestOverlap(B,D,B.name,D.name,out.dir.name,100)
TestOverlap(C,D,C.name,D.name,out.dir.name,100)
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
return(re.out)
}
#' ParserBedFile4PengVenn
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/BedFromPeng/"
#' input.file.pattern="*combine_chr_st_end.bed"
#'
#' input.file.pattern="*combine_chr_st.bed"
#'
#'
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng.4.venn<-ParserBedFile4PengVenn(dir.name,input.file.pattern,out.dir.name)
#'
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4PengVenn<-function(dir.name,input.file.pattern,out.dir.name){
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",7)
print(file.name.2)
#file.name.3<-file.name.2[-6]
re.out<-lapply(file.name.2,function(u){
re=read.table(u,header=FALSE)
re<-as.character(re[,1])
#colnames(re)=c("Count","GeneName")
re
})
sample.name<-sapply(strsplit(names(re.out),split="_peaks_"),"[[",1)
names(re.out)=sample.name
A=re.out[[1]] #10_2470IUPUI_WT_BM_ASXL1
B=re.out[[2]] #10_WT_BM_ASXL1
C=re.out[[3]] #11_2470IUPUI_WT_BM_SMC1
D=re.out[[5]] #13_2470IUPUI_WT_BM_Rad21
A.name=sample.name[1] #10_2470IUPUI_WT_BM_ASXL1
B.name=sample.name[2] #10_WT_BM_ASXL1
C.name=sample.name[3] #11_2470IUPUI_WT_BM_SMC1
D.name=sample.name[5] #13_2470IUPUI_WT_BM_Rad21
#A <- createRandomRegions(nregions=50, length.mean=5000000, length.sd=3000000)
#B <- c(A[1:25], createRandomRegions(nregions=25, length.mean=500000, length.sd=30000))
#class(A)
#class(B)
# TestOverlap <- function(A, B, A.name,B.name,out.dir.name,n_permutations) {
# NumOverLap<-numOverlaps(A, B, count.once=TRUE)
# pt <- overlapPermTest(A=A, B=B, ntimes=n_permutations)
# png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_overlap_test.png"))
# plot(pt)
# dev.off()
#
# lz <- localZScore(pt=pt, A=A, B=B)
# png(filename=paste0(out.dir.name,A.name,B.name,n_permutations,"_localZScore.png"))
# plot(lz)
# dev.off()
# }
#
# TestOverlap(A,B,A.name,B.name,out.dir.name,100)
# TestOverlap(A,C,A.name,C.name,out.dir.name,100)
# TestOverlap(A,D,A.name,D.name,out.dir.name,100)
#
# TestOverlap(B,C,B.name,C.name,out.dir.name,100)
# TestOverlap(B,D,B.name,D.name,out.dir.name,100)
# TestOverlap(C,D,C.name,D.name,out.dir.name,100)
#
# temp.name<-strsplit(names(file.name.2),split="\\.")
#a = readBed(system.file("extdata", "examples/combined_regions.bed",
# package="LOLA"))
venn.plot <- venn.diagram(
x = re.out[c(1,2)],
filename = paste0(out.dir.name,names(re.out)[1],names(re.out)[2],"_overlap_venn.tiff"),
height = 3000,
width = 3500,
resolution = 1000,
col = "black",
lty = "dotted",
lwd = 1,
fill = c("red","blue"),
alpha = 0.50,
label.col = c(rep("white",3)),
cex = 0.5,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","blue"),
cat.cex = 0.5,
cat.pos = 0.5,
cat.dist = 0.05,
cat.fontfamily = "serif"
)
venn.plot <- venn.diagram(
x = re.out[c(1,3,5)],
filename =paste0(out.dir.name,names(re.out)[1],names(re.out)[3],names(re.out)[5],"_overlap_venn.tiff"),
col = "black",
lty = "dotted",
lwd = 2,
fill = c("red", "orange", "blue"),
alpha = 0.50,
label.col = c(rep("white",7)),
cex = 1,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red", "orange", "blue"),
cat.cex = 0.8,
cat.fontfamily = "serif"
)
return(re.out)
}
#' ParserBedFile4PengXls
#'
#' @param dir.name
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' dir.name="/media/H_driver/2016/Yang/MACS/MACS/"
#' input.file.pattern="*.xls"
#'
#' out.dir.name="/media/H_driver/2016/Yang/Results/"
#'
#' re.from.bed.peng.Xls<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' re.from.bed.peng.Xls.2<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' re.from.bed.peng.Xls.3<-ParserBedFile4PengXls(dir.name,input.file.pattern,out.dir.name)
#'
#' save.image(file=paste0(out.dir.name,"re_save_peng.RData"))
#' savehistory(file=paste0(out.dir.name,"re_save_peng.Rhistory"))
#'
ParserBedFile4PengXls<-function(dir.name,input.file.pattern,out.dir.name){
dir.name=reformatPath(dir.name)
out.dir.name=reformatPath(out.dir.name)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",8)
print(file.name.2)
re.out<-lapply(names(file.name.2),function(u){
#print(names(u))
uu=file.name.2[[u]]
re=read.table(uu,header=TRUE)
colnames(re)[c(7,9)]=c("-10*LOG10(pvalue)","FDR(%)")
temp<-re
temp$start<-temp$start-1
temp$end<-temp$end-1
temp$summit<-temp$summit-1
temp
summitPeak<-temp$start+temp$summit
temp2<-temp
temp2$start<-summitPeak-49
temp2$end<-summitPeak+50
temp2$summit<-summitPeak
temp2$length<-temp2$end-temp2$start+1
temp2
rownames(temp2)<-paste0("MACS_peak_",rownames(temp2))
temp3<-toGRanges(temp2)
names(temp3)<-rownames(temp2)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Mmusculus.UCSC.mm10")
genome(temp3.trimmed)<-"mm10"
seq.temp3<-getAllPeakSequence(temp3.trimmed,genome=genome.mm10)
write2FASTA(seq.temp3, paste0(out.dir.name,u,".fa"))
re2<-list(originalPeak=re,down1Peak=temp,aroundSummit100Peak=temp2,GR=temp3)
re2
})
sample.name<-sapply(strsplit(names(file.name.2),split="_peaks_"),"[[",1)
names(re.out)<-sample.name
return(re.out)
}
#' ParserReadFiles
#'
#' @param input.file.dir
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="~/TestBam/"
#' out.file.dir="~/"
#' testR<-ParserReadFiles(input.file.dir)
#'
#'
ParserReadFiles <- function(input.file.dir,input.file.type) {
#For input
dir.name=input.file.dir
dir.name=reformatPath(dir.name)
file.name=file.path(dir.name,dir(dir.name,recursive = TRUE))
file.name.2<-as.list(file.name)
file.name.3<-lapply(1:length(file.name.2),function(u,input.file.type,file.name.2){
tmp=file.name.2
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
n <- length(grep(input.file.type,file_name))
#if(file_ext(file_name)==input.file.type)
if(n == 1) {
re<-file.path(path_name,file_name)
#names(re)[u]<-file_name
}else
{re<-NULL}
re
},input.file.type,file.name.2)
file.name.4<-file.name.3[lapply(file.name.3, length) > 0]
names(file.name.4)=unlist(lapply(1:length(file.name.4),function(u,file.name.4){
tmp=file.name.4
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
file_name
},file.name.4))
#print(file.name.4)
#For output
#output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
#temp=Sys.time()
#temp1=gsub(":","-",temp)
#temp2=gsub(" ","-",temp1)
#temp3=file.path(output.dir.name,"ReadBam_at_",temp2)
#temp3=output.dir.name
#dir.create(temp3)
re2<-list(input=file.name.4)
return(re2)
}
#' ChipSeq:::getGeneBedName("/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny","peaks.csv","/Volumes/Bioinformatics$/2017/DannyNewData/NewRe2Danny")
#'
getGeneBedName <- function(input.file.dir,file.pattern,output.file.dir) {
re<-ParserReadFiles(input.file.dir,file.pattern)
file.name.2<-re$input
re.out<-file.name.2
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
temp3 =output.file.dir
cmd.1 <- lapply(1:length(re.out),function(u,re.out,temp3){
x <- read.csv(re.out[[u]],header = TRUE)
xx <- x[grep("Promoter",x$annotation),]
chr <- which(colnames(xx) == "seqnames")
gs <- which(colnames(xx) == "geneStart")
ge <- which(colnames(xx) == "geneEnd")
file_name = file_path_sans_ext(basename(re.out[[u]]))
write.table(xx[,c(chr,gs,ge)],file=file.path(temp3,paste0(file_name,"_gene.bed")),quote = F,col.names = F,row.names = F,sep="\t")
write.table(unique(xx$SYMBOL),file=file.path(temp3,paste0(file_name,"_gene_name.txt")),quote = F,col.names = F,row.names = F)
ps <- which(colnames(xx) == "start")
pe <- which(colnames(xx) == "end")
write.table(xx[,c(chr,ps,pe)],file=file.path(temp3,paste0(file_name,"_peaks.bed")),quote = F,col.names = F,row.names = F,sep="\t")
},re.out,temp3)
}
#' PeakCallAndAnnotation
#'
#' Call peak for bam files using macs14 and perform peak annotation using ChIPpeakAnno
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param index.file
#' @param output.file.dir
#' @param genome
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.file.dir="/projects/scratch/bbc/Project/Danny_chip/Alignment/BWA/"
#' input.file.pattern="*.bam"
#' index.file=9
#' output.file.dir="/scratch/projects/bbc/aiminy_project/
#' genome="Hs"
#'
#' PeakCallAndAnnotation(input.file.dir,output.file.dir,genome)
#'
PeakCallAndAnnotation <- function(input.file.dir,output.file.dir,genome) {
re<-ParserReadFiles(input.file.dir,"bam",output.file.dir)
file.name.2<-re$input
output.dir.name=re$output
temp3=paste0(output.dir.name,"_PeakCall")
dir.create(temp3)
re.out<-file.name.2
cmd9="macs14 -t "
cmd10="-f BAM -g hs -n "
cmd11=" -m 6,18 --bw=200 -p 0.00001"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd12=paste(cmd9,x,cmd10,paste0(temp3,"/",paste0(x_name,"_hs_1.00e-05_macs142")),cmd11,sep=" ")
print(cmd12)
system(cmd12, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
#AnnotatePeak2(paste0(temp3,"/"),"*macs142_peaks.bed",7,paste0(output.dir.name,"PeakAnnotation_at_",temp2),genome="Hs")
AnnotatePeak3(paste0(temp3,"/"),paste0(output.dir.name,"_PeakAnnotation"),
genome="Hs")
BamFileSortIndexVisualization2(re,genome)
}
BamFileSortIndexVisualization2 <- function(input,genome) {
#library(ChIPpeakAnno)
re<-input
file.name.2<-re$input
output.dir.name=re$output
temp3=paste0(output.dir.name,"_visualization")
dir.create(temp3)
re.out<-file.name.2
cmd1="samtools sort"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd2=paste0(cmd1," ",x," ",paste0(temp3,"/",x_name,"_sorted"))
#print(cmd2)
system(cmd2)
},re.out,temp3)
cmd3="samtools index"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd4=paste0(cmd3," ",paste0(temp3,"/",x_name,"_sorted.bam"))
#print(cmd2)
system(cmd4)
},re.out,temp3)
cmd5="ngs.plot.r -G hg19 -R tss -C"
cmd6="-O"
cmd7="-L 4000"
#cmd3="-L 4000 -RR 1 -CD 1 -CO \\\"blue\\\""
#ngs.plot.r -G hg19 -R tss -C $1 -O $2 -L 4000 -RR 1 -CD 1 -CO "blue"
#file.name.3<-file.name.2[-6]
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd8=paste(cmd5,paste0(temp3,"/",paste0(x_name,"_sorted.bam")),cmd6,paste0(temp3,"/",paste0(x_name,"_sorted")),cmd7,sep=" ")
print(cmd8)
system(cmd8, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
}
#' ProcessUsingCHIPpeakAnno
#'
#' @param input.file.dir
#' @param input.file.pattern
#' @param output.file.dir
#'
#' @return
#' @export
#'
#' @examples
#' input.file.dir="/media/H_driver/2016/Yang/MACS/MACS/"
#' input.file.pattern="*.bed"
#' output.file.dir="/media/H_driver/2016/Yang/MACS/MACS/"
#'
#' ProcessUsingCHIPpeakAnno(input.file.dir,input.file.pattern,output.file.dir)
#'
ProcessUsingCHIPpeakAnno <- function(input.file.dir,input.file.pattern,output.file.dir) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
dir.create(output.dir.name)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",8)
print(file.name.2)
file.name.3<-file.name.2
sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
names(file.name.3)=sample.name
file.name.4 <-file.name.3[-1]
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
annoData[1:2]
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
re<-makeVennDiagram(re.out[c(2,4,1)],NameOfPeaks=c("SMC1A", "RAD21","ASXL1"),totalTest=35000)
#fisher exact test
UseFisher <- function(temp.ct,index.A,index.B,totalN) {
total.peaks=totalN
A=sum(temp.ct[which(temp.ct[,index.A]==1&temp.ct[,index.B]==1),4])
B=sum(temp.ct[which(temp.ct[,index.A]==1&temp.ct[,index.B]==0),4])
C=sum(temp.ct[which(temp.ct[,index.A]==0&temp.ct[,index.B]==1),4])
D=total.peaks-(A+B+C)
ctb<-matrix(c(A,B,C,D),nrow = 2,dimnames =list(c("In", "Not"),c("In", "Not")))
#re<-fisher.test(ctb)
print(ctb)
re.fisher<-fisher.test(ctb, alternative='greater')[c("p.value","estimate")]
re.fisher
}
temp.ct<-ol$venn_cnt
#A vs B
index.A<-grep("SMC1",colnames(temp.ct))
index.B<-grep("Rad21",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.AB=tempRe$p.value
OR.fisher.AB=tempRe$estimate
#A vs C
index.A<-grep("SMC1",colnames(temp.ct))
index.B<-grep("ASXL1",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.AC=tempRe$p.value
OR.fisher.AC=tempRe$estimate
#B vs C
index.A<-grep("Rad21",colnames(temp.ct))
index.B<-grep("ASXL1",colnames(temp.ct))
tempRe<-UseFisher(temp.ct,index.A,index.B,35000)
pVal.fisher.BC=tempRe$p.value
OR.fisher.BC=tempRe$estimate
pVal.fisher.AB
OR.fisher.AB
pVal.fisher.AC
OR.fisher.AC
pVal.fisher.BC
OR.fisher.BC
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
library(TxDb.Hsapiens.UCSC.mm9.knownGene)
aCR<-assignChromosomeRegion(overlaps, nucleotideLevel=FALSE,
precedence=c("Promoters", "immediateDownstream",
"fiveUTRs", "threeUTRs",
"Exons", "Introns"),
TxDb=TxDb.Mmusculus.UCSC.mm9.knownGene)
barplot(aCR$percentage, las=3)
pie1(aCR$percentage,las=3)
dc<-annoGR(TxDb.Mmusculus.UCSC.mm9.knownGene)
seqinfo(dc)
seqlevels(dc)
seqinfo(overlaps)
seqlevels(overlaps)
#GRCm38/mm10
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
seqinfo(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
seqinfo(overlaps)
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
#GRCm38/mm10
dd<-toGRanges(EnsDb.Mmusculus.v79)
seqinfo(dd)
library(ensembldb)
library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
library(org.Mm.eg.db)
overlaps.anno.with.entrez.id <- addGeneIDs(overlaps.anno,"org.Mm.eg.db",IDs2Add = "entrez_id")
write.csv(as.data.frame(unname(overlaps.anno.with.entrez.id)), paste0(out.dir.name,"other_anno.csv"))
pie1(table(overlaps.anno.with.entrez.id$insideFeature))
library("DBI")
over <- getEnrichedGO(overlaps.anno.with.entrez.id, orgAnn="org.Mm.eg.db",
maxP=.05, minGOterm=10,
multiAdjMethod="BH", condense=TRUE)
# over.gene.symbol <- getEnrichedGO(overlaps.anno.with.entrez.id, orgAnn="org.Mm.eg.db",
# feature_id_type="gene_symbol",
# maxP=.05, minGOterm=10,
# multiAdjMethod="BH",condense=TRUE)
head(over[["bp"]][, -c(3, 10)])
library(org.Hs.eg.db)
e2s = toTable(org.Mm.egSYMBOL)
tempDS=over$bp
tempDS2<-data.frame(apply(tempDS,1,function(u,e2c){
#print(u[1])
x=u[11]
tempId<-unlist(strsplit(as.character(x),split=";"))
index<-which(!is.na(match(e2s[,1],tempId)))
index<-match(tempId,e2s[,1])
geneS<-paste(e2s[index,2], collapse=";")
geneS
#print(geneS)
},e2c))
tempDS3<-cbind(tempDS,tempDS2)
colnames(tempDS3)[12]="GeneSymbol"
Draw4GO <- function(tempDS3) {
x=tempDS3
y=x[order(x$pvalue,decreasing = TRUE),]
z=y[1:10,c(1,3,4,5,6,10)]
Function<-z$Definition
negative_log10p=-log10(z$pvalue)
library(ggplot2)
#ggplot(z, aes(x=z$go.id, y=negative_log10p,fill=factor(z$go.id)))+geom_bar(stat="identity")+geom_hline(yintercept = -log10(0.05))+coord_flip()
ggplot(z, aes(go.id,pvalue, fill = go.id)) +
geom_bar(stat="identity")+ scale_x_discrete(labels=z$count.InDataset, limits=factor(z$go.id))+ scale_fill_discrete(breaks = z$go.id,
name="GO term")+theme(legend.text = element_text(colour="black", size = 11, face = "bold"))+
theme(axis.text.x = element_text(angle = 90, hjust = 1))+ggtitle("GO enrichment analysis")+labs(x="Gene Counts",y="-log p-value")
}
write.table(tempDS3,file=paste0(out.dir.name,"BP_.txt"),row.names = FALSE,quote=FALSE,sep="\t")
#anno <- annoGR(EnsDb.Hsapiens.v79)
ree<-annotatePeakInBatch(overlaps,
AnnotationData=dc,
output="nearestBiDirectionalPromoters",
bindingRegion=c(-2000, 500))
ree2 <- addGeneIDs(ree,
"org.Mm.eg.db",
IDs2Add = "entrez_id")
re<-makeVennDiagram(re.out[c(2,4,1)],NameOfPeaks=c("SMC1A", "RAD21","ASXL1"),totalTest=35000)
library(BSgenome.Mmusculus.UCSC.mm9)
upseqs<-Views(Mmusculus,overlaps)
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
mm9.S<-Seqinfo(genome="mm9")
seqinfo(overlaps,force=TRUE) <- Seqinfo(genome="mm9")
seqlevels(mm9.S) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps) <- mm9.S
goodGR <- trim(overlaps)
overlaps.trimmed<-goodGR
seq<-getAllPeakSequence(overlaps.trimmed,genome=Mmusculus)
seqs.mm9 <- getSeq(Mmusculus,overlaps.trimmed)
write2FASTA(seq, paste0(out.dir.name,"WT_triple.fa"))
## We can also try simulation data
seq.sim.motif <- list(c("t", "g", "c", "a", "t", "g"),
c("g", "c", "a", "t", "g", "c"))
set.seed(1)
seq.sim <- sapply(sample(c(2, 1, 0), 1000, replace=TRUE, prob=c(0.07, 0.1, 0.83)),
function(x){
s <- sample(c("a", "c", "g", "t"),
sample(100:1000, 1), replace=TRUE)
if(x>0){
si <- sample.int(length(s), 1)
if(si>length(s)-6) si <- length(s)-6
s[si:(si+5)] <- seq.sim.motif[[x]]
}
paste(s, collapse="")
})
os <- oligoSummary(seq, oligoLength=6, MarkovOrder=3,
quickMotif=TRUE)
zscore <- sort(os$zscore, decreasing=TRUE)
h <- hist(zscore, breaks=100, main="Histogram of Z-score")
text(zscore[1:2], rep(5, 2),
labels=names(zscore[1:2]), adj=0, srt=90)
pfms <- mapply(function(.ele, id)
new("pfm", mat=.ele, name=paste("SAMPLE motif", id)),
os$motifs, 1:length(os$motifs))
motifStack(pfms[[1]])
motifStack(pfms[[2]])
motifStack(pfms[[3]])
motifStack(pfms[[4]])
}
# input.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
# input.file.pattern="*macs142_peaks.bed"
# output.file.dir="/media/H_driver/2016/Danny/Danny_chip_PeakCall/"
#
# AnnotatePeak(input.file.dir,input.file.pattern,8,output.file.dir,genome="Mm")
# AnnotatePeak(input.file.dir,input.file.pattern,7,output.file.dir,genome="Hs")
AnnotatePeak <- function(input.file.dir,input.file.pattern,index.file,output.file.dir,genome) {
library(ChIPpeakAnno)
dir.name=input.file.dir
input.file.pattern=input.file.pattern
dir.name=reformatPath(dir.name)
output.dir.name=reformatPath(output.file.dir)
#print(output.dir.name)
temp=Sys.time()
temp1=gsub(":","-",Sys.time())
temp2=gsub(" ","-",temp1)
temp3=paste0(output.dir.name,"AnalysisResults_at_",temp2)
dir.create(temp3)
file.name=paste0(dir.name,dir(dir.name,recursive = TRUE,pattern=input.file.pattern))
file.name.2<-as.list(file.name)
names(file.name.2)=sapply(strsplit(file.name,split="\\/"),"[[",index.file)
print(file.name.2)
file.name.3<-file.name.2
#sample.name<-sapply(strsplit(names(file.name.3),split="_peaks_"),"[[",1)
#names(file.name.3)=sample.name
file.name.4 <-file.name.3[-1]
re.out<-lapply(file.name.4,function(u){
re=toGRanges(u,format="BED")
#colnames(re)=c("Count","GeneName")
re
})
head(re.out[[1]])
re.out.L<-lapply(re.out,function(u){
re=length(u)
#colnames(re)=c("Count","GeneName")
re
})
if(genome=="Mm"){
annoData <- toGRanges(EnsDb.Mmusculus.v75, feature="gene")
ol <- findOverlapsOfPeaks(re.out[c(2,4,1)])
overlaps<-ol$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed///10_WT_BM_ASXL1_peaks.bed`
binOverFeature(overlaps, annotationData=annoData,
radius=5000, nbins=20, FUN=length, errFun=0,
ylab="count",
main="Distribution of aggregated peak numbers around TSS")
overlaps.trimmed<-trim(overlaps, use.names=TRUE)
library(EnsDb.Mmusculus.v79)
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
#seqinfo(dd.GRCm39.mm10)
#seqlevels(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
"chr14","chr15","chr16","chr17","chr18","chr19","chr2",
"chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
seqinfo(overlaps)<-seqinfo(dd.GRCm39.mm10)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(overlaps.trimmed) <- seqlevelsStyle(dd.GRCm39.mm10)
overlaps.anno<-annoPeaks(overlaps.trimmed,dd.GRCm39.mm10)
write.table(overlaps.anno,file=paste0(temp3,"/","annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
}else if(genome=="Hs"){
library(EnsDb.Hsapiens.v75)
#annoData<-toGRanges(EnsDb.Hsapiens.v75, feature="gene")
dd.hs<-toGRanges(EnsDb.Hsapiens.v75)
print(seqinfo(dd.hs))
print(seqlevels(dd.hs))
#print(seqlevels(dd.hs)[,1])
#print(seqlevels(re.out[[1]])[,1])
# seqlevels(dd.hs,force=TRUE) <- c("chr1","chr10","chr11","chr12","chr13",
# "chr14","chr15","chr16","chr17","chr18","chr19","chr2",
# "chr3","chr4","chr5","chr6","chr7","chr8","chr9","chrX","chrY")
#temp4=
re.out.L<-lapply(1:length(re.out),function(u,re.out,dd.hs){
x=re.out[[u]]
x_name=names(re.out)[u]
seqlevels(dd.hs,force=TRUE)<-seqinfo(x)@seqnames
#print(seqinfo(re.out.trimmed))
#print(seqlevels(re.out.trimmed))
seqinfo(x)<-seqinfo(dd.hs)
#GRCm38/mm10
#dd<-toGRanges(EnsDb.Mmusculus.v79)
#seqinfo(dd)
#library(ensembldb)
#library(GenomeInfoDb)
seqlevelsStyle(x) <- seqlevelsStyle(dd.hs)
re.out.trimmed<-trim(x, use.names=TRUE)
overlaps.anno<-annoPeaks(re.out.trimmed,dd.hs)
write.table(overlaps.anno,file=paste0(temp3,"/",x_name,"_annotation.txt"),row.names = FALSE,quote=FALSE,sep="\t")
},re.out,dd.hs)
}
}
ProcessUsingLOLA <- function() {
#dbPath = system.file("extdata", "hg19", package="LOLA")
dbPath = "/media/H_driver/2016/Yang/Results/data/groups/lab_bock/shared/resources/regions/LOLACore/mm10"
regionDB = loadRegionDB(dbLocation=dbPath)
#read triple overlap peaks
regionSetA = readBed("/media/H_driver/2016/Yang/Results/WT_triple_overlap.bed")
#read each peak
regionSet.WT.ASXL1=readBed("/media/H_driver/2016/Yang/BedFromPeng/10_WT_BM_ASXL1_peaks_from_PeakCall4Yang.bed")
regionSet.WT.SMC1=readBed("/media/H_driver/2016/Yang/BedFromPeng/11_2470IUPUI_WT_BM_SMC1_peaks_from_PeakCall4Yang.bed")
regionSet.WT.Rad21=readBed("/media/H_driver/2016/Yang/BedFromPeng/13_2470IUPUI_WT_BM_Rad21_peaks_from_PeakCall4Yang.bed")
db.841<-regionDB$regionGRL
userSets = GRangesList(regionSet.WT.ASXL1,regionSet.WT.SMC1,regionSet.WT.Rad21)
db.841.2<-c(db.841,userSets)
userUniverse<-buildRestrictedUniverse(userSets)
userUniverse.844<-buildRestrictedUniverse(db.841.2)
re.SMC1A.RAD21.only<-makeVennDiagram(re.out[c(2,4)],NameOfPeaks=c("SMC1A", "RAD21"),totalTest=35000)
ol.SMC1A.RAD21.only <- findOverlapsOfPeaks(re.out[c(2,4)])
ol.SMC1A.RAD21.only.2<-ol.SMC1A.RAD21.only$peaklist$`11_2470IUPUI_WT_BM_SMC1_peaks.bed///13_2470IUPUI_WT_BM_Rad21_peaks.bed`
checkUniverseAppropriateness(ol.SMC1A.RAD21.only.2,userUniverse.844, cores = 1,fast = FALSE)
locResults2 = runLOLA(userSets,userUniverse,regionDB, cores=1)
locResults = runLOLA(regionSetA,userUniverse,regionDB, cores=1)
#SMC1A and RAD21 overlap
locResults.SMC1A.RAD21 = runLOLA(ol.SMC1A.RAD21.only.2,userUniverse,regionDB, cores=4)
head(locResults.SMC1A.RAD21)
pValue<-as.data.frame(10^(-locResults.SMC1A.RAD21$pValueLog))
temp<-as.data.frame(locResults.SMC1A.RAD21)
locResults.SMC1A.RAD21.with.p<-cbind(temp[,1:3],pValue,temp[,4:23])
colnames(locResults.SMC1A.RAD21.with.p)[4]="pValue"
#It takes extensive computation resource, be carefully to run
locResults.SMC1A.RAD21.based.large.universe = runLOLA(ol.SMC1A.RAD21.only.2,userUniverse.844,regionDB, cores=4)
write.table(locResults.SMC1A.RAD21.with.p,
file=paste0(out.dir.name,"SMC1A.RAD21.with.Other.Antibody.csv"),sep=",",
quote = FALSE,row.names = FALSE,col.names = TRUE)
write.table(locResults,file=paste0(out.dir.name,"Other.csv"),sep=",",quote = FALSE,row.names = FALSE,col.names = TRUE)
listRegionSets(regionDB)
extractEnrichmentOverlaps(locResults,regionSetA, regionDB)
loadCaches("/media/H_driver/2016/Yang/Results/LOLACoreCaches_latest.tgz")
save.image(file =paste0(out.dir.name,"re_search.RData"))
# regionSetB = readBed("lola_vignette_data/setB_100.bed")
# regionSetC = readBed("lola_vignette_data/setC_100.bed")
# activeDHS = readBed("lola_vignette_data/activeDHS_universe.bed")
# data("sample_universe", package="LOLA")
#
# dbPath = system.file("extdata", "hg19", package="LOLA")
# regionDB = loadRegionDB(dbLocation=dbPath)
# data("sample_universe", package="LOLA")
# data("sample_input", package="LOLA")
#
# getRegionSet(regionDB, collections="ucsc_example", filenames="vistaEnhancers.bed")
# getRegionSet(dbPath, collections="ucsc_example", filenames="vistaEnhancers.bed")
#
# res = runLOLA(userSets, userUniverse, regionDB, cores=1)
# locResult = res[2,]
}
#' parser4diffbind
#'
#' parser4diffbind
#'
#' @param input.sample.file
#' @param input.bed.dir
#' @param input.file.pattern
#'
#' @return
#' @export
#'
#' @examples
#'
#' input.sample.file = "/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv"
#' input.bed.dir = "/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/"
#' input.file.pattern = "bed"
#' output.dir="BindDiff_4_7_2017-cutoff_10k_test"
#' output.dir="/Volumes/Bioinformatics$/2017/DannyNewData/testRun"
#' resmcf<-parser4diffbind(input.sample.file,input.bed.dir,input.file.pattern,output.dir)
#'
parser4diffbind<-function(input.sample.file,input.bed.dir,input.file.pattern,output.dir,
summit_peak=NULL){
sample.info<-read.csv(input.sample.file,header = TRUE)
sample.info.2 <- sample.info[,-dim(sample.info)[2]]
out.dir.name = dirname(input.sample.file)
re<-ParserReadFiles(input.bed.dir,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2<-FL(re.bed,'peaks')
if(!is.null(summit_peak)){
re.summits.only.bed<-FL(re.bed,'summits')
}
#put peak files into database
re.peaks.only.bed.3<-list_to_df(re.peaks.only.bed.2)
print(re.peaks.only.bed.3)
pos <- regexpr('R1', re.peaks.only.bed.3$ID)
ID2 <- substr(re.peaks.only.bed.3$ID,1,pos+1)
re.peaks.only.bed.3 <- cbind(ID2,re.peaks.only.bed.3)
colnames(re.peaks.only.bed.3)=c("ID","ID2")
re.peaks.only.bed.4<-merge(sample.info.2,re.peaks.only.bed.3,by="ID")
replicate<-apply(re.peaks.only.bed.4,1,function(x){
ID=as.character(x[1])
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
p1=nchar(ID)+2
p2=regexpr(TF,ID2)
p2=p2-2
xx=substr(ID2,p1,p2)
cell=substr(xx,1,nchar(xx)-2)
rep=substr(xx,nchar(xx),nchar(xx))
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
z
})
res <- do.call(rbind.data.frame,replicate)
colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
re.peaks.only.bed.5 <- res
mcf<-NULL
for(i in 1:dim(re.peaks.only.bed.5)[1]){
#peaks.v=readBed(as.character(re.peaks.only.bed.5[i,6]))
peaks.v <- readPeakFile(as.character(re.peaks.only.bed.5[i,6]), as = "GRanges")
peak.caller.v="macs142"
sampID.v=as.character(re.peaks.only.bed.5[i,1])
sampID.v=substr(sampID.v,1,nchar(sampID.v)-5)
tissue.v=as.character(re.peaks.only.bed.5[i,2])
factor.v=as.character(re.peaks.only.bed.5[i,3])
condition.v=as.character(re.peaks.only.bed.5[i,4])
replicate.v=as.numeric(as.character(re.peaks.only.bed.5[i,5]))
if(is.null(mcf)){
mcf <- dba.peakset(NULL,peaks=peaks.v,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}else{
mcf <- dba.peakset(mcf,peaks=peaks.v,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}
}
print(mcf)
temp3=output.dir
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
save(mcf,file=file.path(temp3,"mcf.RData"))
pdf(file.path(temp3,"corrmap.pdf"))
dba.plotHeatmap(mcf,margin=23)
dev.off()
GetResultsFromDiffBind(mcf,"yes",temp3)
return(mcf)
}
#' Title
#'
#' @param dir.name
#'
#' @return
#' @export
#'
#' @examples
#' dir.name="/media/H_driver/2016/Yang/MACS/MACS/"
#' reformatPath(dir.name)
reformatPath <- function(dir.name){
CheckOPS=Sys.info()[['sysname']]
if(CheckOPS=="Darwin"){
if(length(grep('H_driver',dir.name))!=0){
temp=unlist(strsplit(dir.name,split="\\/"))
temp[2]="Volumes"
temp[3]="Bioinformatics$"
dir.name=paste0(paste0(temp,collapse = "/"),"/")
}
}
return(dir.name)
}
#' useHOMER
#'
#' @return
#' @export
#'
#' @examples
#' res <- useHOMER("findMotifs.pl")
#' system(res)
#'
useHOMER <- function(function.name){
R_LIB=.libPaths()
PATH1=Sys.getenv("PATH")
homer_Lib=file.path(R_LIB,"ChipSeq/homer/bin")
Sys.setenv(PATH=paste0(homer_Lib,":",PATH1))
cmd1 <- Sys.which(function.name)[[1]]
PATH2=Sys.getenv("PATH")
cat(PATH1,"\n")
cat(PATH2,"\n")
return(cmd1)
}
# Filtering list based on certain pattern of the names of list elements
FL <- function(re.bed, patt) {
re.peaks.only.bed<-lapply(1:length(re.bed),function(u,re.bed,patt){
tmp=re.bed
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
#pos_summits=regexpr('summits',file_name)
pos_peaks=regexpr(patt,file_name)
if(pos_peaks>0){
y<-x
}else{
y<-NULL
}
y
},re.bed,patt)
re.peaks.only.bed.2<-re.peaks.only.bed[lapply(re.peaks.only.bed,length) > 0]
names(re.peaks.only.bed.2)=unlist(lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2){
tmp=re.peaks.only.bed.2
x=tmp[[u]]
path_name=dirname(x)
file_name=basename(x)
file_name
},re.peaks.only.bed.2))
return(re.peaks.only.bed.2)
}
list_to_df <- function(list_for_df) {
list_for_df <- as.list(list_for_df)
nm <- names(list_for_df)
if (is.null(nm))
nm <- seq_along(list_for_df)
ID<-sapply(1:length(nm),function(u,nm){
x=nm[u]
pos=regexpr("\\.",x)
pos1=pos-1
y<-substr(x,1,pos1)
y
},nm)
df <- data.frame(ID=ID,name = nm, stringsAsFactors = FALSE)
df$value <- unname(list_for_df)
df
}
get_os <- function(){
sysinf <- Sys.info()
if (!is.null(sysinf)){
os <- sysinf['sysname']
if (os == 'Darwin')
os <- "osx"
} else { ## mystery machine
os <- .Platform$OS.type
if (grepl("^darwin", R.version$os))
os <- "osx"
if (grepl("linux-gnu", R.version$os))
os <- "linux"
}
tolower(os)
}
#' plotBam2
#'
#' @export
#' @example
#'
#' input.sample.file <- '/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny.csv'
#'
#' input.bam.file <- '/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny/Bam_files.txt'
#'
#' output.dir <- '/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny/BamMerged'
#'
#' re <- plotBam2(input.sample.file,input.bam.file,output.dir)
#'
#'
plotBam2 <- function(input.sample.file,input.bam.file,output.dir)
{
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$re11
cell.name <- unique(unlist(lapply(cellInfo$Type_Cell,function(u){substr(as.character(u),1,nchar(as.character(u))-2)})))
cmd="samtools merge"
cmd.3 <- lapply(cell.name,function(u,cellInfo,output.dir){
cat(u,"\n")
x <- cellInfo[which(substr(as.character(cellInfo$Type_Cell),1,nchar(as.character(cellInfo$Type_Cell))-2)==u),]
x$Cell_TF <- gsub(" ","-",x$Cell_TF)
tf <- as.character(x$Type_TF)
tf.unique <- unique(tf)
y <- combn(1:length(tf),2,function(x)list(c(x[1],x[2])))
yy <- lapply(tf.unique,function(u,tf,y){
x <- which(tf %in% u)
x
},tf,y)
yyy <- setdiff(y,yy)
yyyy <- yyy[lapply(yyy, length) > 0]
cmd.2 <- lapply(yyyy,function(u,x,output.dir){
cmd.1 <- paste(cmd,file.path(output.dir,paste0(paste(x[c(u),]$Cell_TF,collapse ="-"),"_merged.bam")),paste(x[c(u),]$file.name,collapse = " "),collapse = " ")
},x,output.dir)
cmd.2
},cellInfo,output.dir)
cmd.4 <- unlist(cmd.3)
mergeBamFilesUseJobArray(cmd.4)
}
mergeBamFilesUseJobArray <- function(cmd)
{
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
total <- system('echo $LSB_JOBINDEX_END',intern = TRUE)
cat(index,"\n\n")
cat(total,"\n\n")
u <- as.integer(index)
system(cmd[u],intern = TRUE)
cat(cmd,"\n\n")
}
#' R -e 'library(DoGs);library(ChipSeq);ChipSeq:::runPlotBam2("~/Danny/SampleID_INFO_ChIP_new_Danny.csv","~/Danny/Alignment/BWA/bam_files.txt","/scratch/projects/bbc/aiminy_project/DannyBamMerged")'
runPlotBam2 <-
function(input.sample.file,
input.bam.file,
output.dir) {
# This setting works
Rfun1 <-
'library(DoGs);library(ChipSeq);re <- ChipSeq:::plotBam2('
input1 <- input.sample.file
input2 <- input.bam.file
output <- output.dir
Rinput <-
paste0('\\"',
input1,
'\\",',
'\\"',
input2,
'\\",',
'\\"',
output,
'\\"')
Rfun2 <- ')'
Rfun3 <- paste0(Rfun1, Rinput, Rfun2)
n.job <- 12
mergeBam <-
DoGs:::createBsubJobArrayRfun(Rfun3, paste0("mergeBam[1-", n.job, "]"), NULL)
system(mergeBam)
}
#' bsub -P bbc -J "bamPlot" -o %J.bamPlot.log -e %J.bamPlot.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::plotMergedBamWgene("/scratch/projects/bbc/aiminy_project/DannyNewNgsPlot4Merged","*_sorted.bam","~/all_common_gene_unique.txt","NgsPlot4Merged")'
#'
plotMergedBamWgene <-
function(input.bam.file.dir,
file.pattern,
input.gene.list,
output.dir,
ngs.para = c("hg19", 4000, 1, 1, "total"),
add.input = NULL)
{
cellInfo <-
list.files(
input.bam.file.dir,
pattern = file.pattern,
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE
)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
cellInfo.run <- lapply(1:length(cellInfo), function(u, cellInfo,
temp3)
{
xx <- cellInfo[u]
xxx <- file_path_sans_ext(basename(xx))
cmd12 = paste(xx, input.gene.list, xxx, sep = "\t")
cat(cmd12, file = file.path(temp3, paste0(xxx, "_config_cJunAndp27.txt")), sep = "\t")
}, cellInfo, temp3)
# dir.name=temp3 dir.name=reformatPath(dir.name)
file.name <-
list.files(
temp3,
pattern = "*_config_cJunAndp27.txt",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE
)
file.name.2 <- as.list(file.name)
zzz <- unlist(file.name.2)
lapply(1:length(zzz), function(u, zzz)
{
dir.name = dirname(zzz[u][[1]])
file_name = file_path_sans_ext(basename(zzz[u][[1]]))
cmd = paste(
"ngs.plot.r -G hg19 -R tss -C",
zzz[u][[1]],
"-O",
file.path(dir.name,
paste0(file_name, ".tss")),
"-T",
file_name,
"-L 4000 -RR 1 -CD 1 -GO total",
sep = " "
)
cmd2 = cmd
cat(cmd2, "\n")
cat("\n")
system(cmd2)
}, zzz)
return(re)
}
#mcf.with.zhao <- ChipSeq:::addMoreBed2mcf("/Volumes/Bioinformatics$/2017/DannyNewData/SampleID_INFO_ChIP_new_Danny_zhao.csv","~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny","/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall","bed$",mcf=NULL,"/Volumes/Bioinformatics$/2017/DannyNewData/testRun5")
addMoreBed2mcf<-function(input.sample.file,input.bam.dir,input.bed.dir,input.file.pattern,mcf=NULL,output.dir){
bam.file <- prepareBamFile(input.bam.dir)
sample.info<-read.csv(input.sample.file,header = TRUE)
sample.info.2 <- sample.info[,-dim(sample.info)[2]]
#print(sample.info.2)
#out.dir.name = dirname(input.sample.file)
re<-ParserReadFiles(input.bed.dir,input.file.pattern)
re.bed<-re$input
#print(re.bed)
re.peaks.only.bed.2<-Filter(function(x) !any(grepl("summits", x)), re.bed)
# if(!is.null(summit_peak)){
# re.summits.only.bed<-FL(re.bed,'summits')
# }
#put peak files into database
re.peaks.only.bed.3<-list_to_df(re.peaks.only.bed.2)
print(re.peaks.only.bed.3)
ll <- re.peaks.only.bed.3$ID
ID2 <- unlist(lapply(ll,function(u) {
pos <- regexpr('R1',u)
if(pos>0){
id <- substr(u,1,pos+1)
}else{
id <- u
}
id
}))
# if(u>0){
# ID2 <- substr(ll,1,pos+1)
# }else{
# ID2 <- ll
# }
# pos <- regexpr('R1', re.peaks.only.bed.3$ID)
#
# print(pos)
#
# ll <- re.peaks.only.bed.3$ID
#
# ID2 <- lapply(pos,function(u,ll) {
#
# if(u>0){
# ID2 <- substr(ll,1,pos+1)
# }else{
# ID2 <- ll
# }
#
# },ll)
re.peaks.only.bed.3 <- cbind(ID2,re.peaks.only.bed.3)
#print(re.peaks.only.bed.3)
colnames(re.peaks.only.bed.3)=c("ID","ID2")
#
re.peaks.only.bed.4<-merge(sample.info.2,re.peaks.only.bed.3,by="ID")
#
cat("4\n")
print(re.peaks.only.bed.4)
cat("\n")
re.peaks.only.bed.with.bam<-merge(bam.file,re.peaks.only.bed.4,by="ID")
print(re.peaks.only.bed.with.bam)
colnames(re.peaks.only.bed.with.bam)[6]="SampleID"
replicate<-apply(re.peaks.only.bed.4,1,function(x){
# pos <- regexpr('R1', re.peaks.only.bed.3$ID)
ID=as.character(x[1])
pos <- regexpr('R1',ID)
if(pos>0){
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
p1=nchar(ID)+2
p2=regexpr(TF,ID2)
p2=p2-2
xx=substr(ID2,p1,p2)
cell=substr(xx,1,nchar(xx)-2)
rep=substr(xx,nchar(xx),nchar(xx))
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
}else
{
cell=as.character(x[2])
TF=as.character(x[3])
ID2=as.character(x[4])
#p1=nchar(ID)+2
#p2=regexpr(TF,ID2)
#p2=p2-2
#xx=substr(ID2,p1,p2)
#cell=substr(xx,1,nchar(xx)-2)
rep=1
file=as.character(x[6])
condition=paste(cell,TF,sep=":")
}
z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
z
})
res <- do.call(rbind.data.frame,replicate)
colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
colnames(res)[1] = "SampleID"
sample.4.dba<-merge(res,re.peaks.only.bed.with.bam,by="SampleID")
re.peaks.only.bed.5 <- res
# replicate<-apply(re.peaks.only.bed.4,1,function(x){
# ID=as.character(x[1])
# cell=as.character(x[2])
# TF=as.character(x[3])
# ID2=as.character(x[4])
# p1=nchar(ID)+2
# p2=regexpr(TF,ID2)
# p2=p2-2
# xx=substr(ID2,p1,p2)
# cell=substr(xx,1,nchar(xx)-2)
# #rep=1
# file=as.character(x[6])
# condition=paste(cell,TF,sep=":")
# z <- data.frame(t(c(ID2,cell,TF,condition,rep,file)))
# z
# })
#
# res <- do.call(rbind.data.frame,replicate)
# colnames(res) <- c("ID","Cell","Ab","Cell_Ab","Replicate","File")
#
#
# re.peaks.only.bed.5 <- res
print(re.peaks.only.bed.5)
mcf<-mcf
ll <- list()
for(i in 1:dim(re.peaks.only.bed.5)[1]){
#peaks.v=readBed(as.character(re.peaks.only.bed.5[i,6]))
peaks.v <- readPeakFile(as.character(re.peaks.only.bed.5[i,6]), as = "GRanges")
peaks.v.sorted <- DiffBind:::pv.peaksort(as.data.frame(peaks.v))
#peaks.v.sorted <- peaks.v
##print(peaks.v)
ll[[i]] <- peaks.v.sorted
peak.caller.v="macs142"
sampID.v=as.character(re.peaks.only.bed.5[i,1])
pos <- regexpr('R1',sampID.v)
if(pos>0){
sampID.v=substr(sampID.v,1,nchar(sampID.v)-5)
}else{
sampID.v = sampID.v
}
tissue.v=as.character(re.peaks.only.bed.5[i,2])
factor.v=as.character(re.peaks.only.bed.5[i,3])
condition.v=as.character(re.peaks.only.bed.5[i,4])
replicate.v=as.numeric(as.character(re.peaks.only.bed.5[i,5]))
if(is.null(mcf)){
mcf <- dba.peakset(NULL,peaks=peaks.v.sorted,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}else{
mcf <- dba.peakset(mcf,peaks=peaks.v.sorted,peak.caller=peak.caller.v, sampID=sampID.v,tissue=tissue.v,
factor=factor.v,condition=condition.v,replicate=replicate.v)
}
}
print(mcf)
temp3=output.dir
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
save(mcf,file=file.path(temp3,"mcf.RData"))
pdf(file.path(temp3,"corrmap.pdf"))
dba.plotHeatmap(mcf,margin=23)
dev.off()
# #GetResultsFromDiffBind(mcf,"yes",temp3)
re <- list(mcf=mcf,ll = ll,re.peaks.only.bed.5 = re.peaks.only.bed.5,re.peaks.only.bed.with.bam = re.peaks.only.bed.with.bam, sample.4.dba = sample.4.dba)
return(re)
}
#ChipSeq:::GetResultsFromDiffBind2(temp,"/Volumes/Bioinformatics$/2017/DannyNewData/testRun2")
GetResultsFromDiffBind2<-function(mcf7,output.file.dir){
temp<-mcf.with.zhao
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
#if(Mergereplicates=="yes"){
temp22<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp22)
##need to set the flexiable number identified from data sets
# t <- temp2
# temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
# }else{
# temp22<-temp
# }
temp22<-dba(temp22,mask =17:28)
print(temp22)
po1 <- c(1,2,6,10)
po2 <- c(1,2)
temp3=file.path(output.file.dir,"Venn2")
if(!dir.exists(temp3))
{
dir.create(temp3)
}
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po1,main=paste0(colnames(temp22$class)[po1],collapse = "-vs-"))
dev.off()
png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
dba.plotVenn(temp22,mask=po2,main=paste0(colnames(temp22$class)[po2],collapse = "-vs-"))
dev.off()
# Tissue1<-temp22$class[2,]
# Tissue2<-unique(temp22$class[2,])
#
# TF<-unique(temp22$class[3,])
# TF.n<-length(TF)
#
# temp3=file.path(output.file.dir,"Venn")
#
# if(!dir.exists(temp3))
# {
# dir.create(temp3)
# }
#
# for(i in 1:length(Tissue2)){
# po<-which(Tissue1 %in% Tissue2[i])
#
# print(po)
#
# if(length(po)==2)
# {
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po,main=Tissue2[i])
# dev.off()
#
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
# dev.off()
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
# dev.off()
#
# }else
# {
# cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
# }
# }
#
# p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
#
# po<-which(Tissue1 %in% Tissue2[u])
#
# if(length(po)==2)
# {
# common.peaks<-dba.overlap(temp22,mask=po)
# y<-common.peaks$inAll
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# common.peaks.1<-dba.overlap(temp22,mask=po1)
# y1<-common.peaks.1$inAll
#
# common.peaks.2<-dba.overlap(temp22,mask=po2)
# y2<-common.peaks.2$inAll
#
# y<-list(y1=y1,y2=y2)
#
# names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
# names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
#
# }else{
# y<-NULL}
# y
# },Tissue1,Tissue2,temp22)
#
# names(p.common)<-Tissue2
#
# p.common<-unlist(p.common,recursive = F)
#
# p.common<-p.common[lapply(p.common,length) > 0]
#
# if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
# {
# dir.create(file.path(output.file.dir,"common_peaks_bed"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
# }
#
# #output common peaks to bed files
#
# lapply(1:length(p.common),function(u,p.common,output.file.dir){
# x=p.common[[u]]
#
# x_name=names(p.common)[u]
#
# df <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=strand(x))
#
# #assign strand
# df.str <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=c(rep(".", length(x))))
#
# df.str.1<-df.str[-grep("random",df.str$seqnames),]
#
# df.str.2<-df.str.1
#
# df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
# col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# },p.common,output.file.dir)
#
# AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
# ,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
#
# return(p.common)
}
useChIPpeakAnno <- function(mcf.with.zhao) {
#findOverlapsOfPeaks(mcf.with.zhao$ll[c(17,11,18)])
overlap <- findOverlapsOfPeaks(mcf.with.zhao$ll[17][[1]],mcf.with.zhao$ll[11][[1]],mcf.with.zhao$ll[18][[1]])
bed.li<- overlap$overlappingPeaks[2][[1]][,2:4]
write.table(bed.li,file="cJun_Li.bed",sep="\t",row.names = FALSE,
col.names = FALSE,quote=FALSE)
bed.zhao <- overlap$overlappingPeaks[2][[1]][,8:10]
write.table(bed.zhao,file="cJun_zhao.bed",sep="\t",row.names = FALSE,
col.names = FALSE,quote=FALSE)
s1 <- makeVennDiagram(mcf.with.zhao$ll[c(17,11,18)], NameOfPeaks=c("cJun_Li","cJun_1833","cJun_zhao"),
totalTest=20000,scaled=F, euler.d=F,fill = c("red","green","blue"),
alpha = 0.50,
label.col = c(rep("black",7)),
cex = 2,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","green","blue"),connectedPeaks = "keepAll")
s2 <- makeVennDiagram(mcf.with.zhao$ll[c(17,3,18)], NameOfPeaks=c("cJun_Li","cJun_231","cJun_zhao"),
totalTest=20000,scaled=F, euler.d=F,fill = c("red","green","blue"),
alpha = 0.50,
label.col = c(rep("black",7)),
cex = 2,
fontfamily = "serif",
fontface = "bold",
cat.col = c("red","green","blue"),connectedPeaks = "keepAll")
}
# SampleID,Tissue,Factor,Condition,Treatment,Replicate,bamReads,ControlID,bamControl,Peaks,PeakCaller
# BT4741,BT474,ER,Resistant,Full-Media,1,reads/Chr18_BT474_ER_1.bam,BT474c,reads/Chr18_BT474_input.bam,peaks/BT474_ER_1.bed.gz,bed
# BT4742,BT474,ER,Resistant,Full-Media,2,reads/Chr18_BT474_ER_2.bam,BT474c,reads/Chr18_BT474_input.bam,peaks/BT474_ER_2.bed.gz,bed
# MCF71,MCF7,ER,Responsive,Full-Media,1,reads/Chr18_MCF7_ER_1.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_1.bed.gz,bed
# MCF72,MCF7,ER,Responsive,Full-Media,2,reads/Chr18_MCF7_ER_2.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_2.bed.gz,bed
# MCF73,MCF7,ER,Responsive,Full-Media,3,reads/Chr18_MCF7_ER_3.bam,MCF7c,reads/Chr18_MCF7_input.bam,peaks/MCF7_ER_3.bed.gz,bed
# T47D1,T47D,ER,Responsive,Full-Media,1,reads/Chr18_T47D_ER_1.bam,T47Dc,reads/Chr18_T47D_input.bam,peaks/T47D_ER_1.bed.gz,bed
# T47D2,T47D,ER,Responsive,Full-Media,2,reads/Chr18_T47D_ER_2.bam,T47Dc,reads/Chr18_T47D_input.bam,peaks/T47D_ER_2.bed.gz,bed
# MCF7r1,MCF7,ER,Resistant,Full-Media,1,reads/Chr18_TAMR_ER_1.bam,TAMRc,reads/Chr18_TAMR_input.bam,peaks/TAMR_ER_1.bed.gz,bed
# MCF7r2,MCF7,ER,Resistant,Full-Media,2,reads/Chr18_TAMR_ER_2.bam,TAMRc,reads/Chr18_TAMR_input.bam,peaks/TAMR_ER_2.bed.gz,bed
# ZR751,ZR75,ER,Responsive,Full-Media,1,reads/Chr18_ZR75_ER_1.bam,ZR75c,reads/Chr18_ZR75_input.bam,peaks/ZR75_ER_1.bed.gz,bed
# ZR752,ZR75,ER,Responsive,Full-Media,2,reads/Chr18_ZR75_ER_2.bam,ZR75c,reads/Chr18_ZR75_input.bam,peaks/ZR75_ER_2.bed.gz,bed
#prepareBamFile("~/Dropbox (BBSR)/BWA")
prepareBamFile <- function(input.bam.dir,file.pattern){
bam <- list.files(path = input.bam.dir, pattern=file.pattern, all.files = TRUE,
full.names = TRUE, recursive = TRUE,
ignore.case = FALSE, include.dirs = TRUE, no.. = FALSE)
bam2 <- as.data.frame(cbind(basename(bam),bam))
ID <- unlist(lapply(bam2[,1],function(u)
{
y <- u
pos <- regexpr("\\.", y)
pos <- pos - 1
y <- substr(y, 1, pos)
y
}))
bam3 <- cbind(ID,bam2)
bam3
}
# tamoxifen <- prepareSampleSheet(mcf.with.zhao,"~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/BamDanny")
prepareSampleSheet <- function(mcf.with.zhao,output.file.dir){
sampleSheet <- mcf.with.zhao$sample.4.dba[,c(1,2,3,5,9,13)]
sampleSheet2 <- cbind(sampleSheet[,1:3],rep("Danny-Treatment",dim(sampleSheet)[1]),rep("Danny-condition",dim(sampleSheet)[1]),sampleSheet[,4:6],rep("bed",dim(sampleSheet)[1]))
colnames(sampleSheet2)=c("SampleID","Tissue","Factor","Condition","Treatment","Replicate","bamReads","Peaks","PeakCaller")
fwrite(sampleSheet2, file ="sampleSheet2.csv")
tamoxifen <- dba(sampleSheet="sampleSheet2.csv")
tamoxifen
tamoxifen <- dba.count(tamoxifen, summits=250)
tamoxifen <- dba.contrast(tamoxifen, group1 = c(1,2),group2=c(3,4),name1="cJun_231_DD", name2="cJun_231")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(1,2),group2=c(11,12),name1="cJun_231_DD", name2="cJun_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(3,4),group2=c(11,12),name1="cJun_231", name2="cJun_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(13,14),group2=c(5,6),name1="p27_231_DD", name2="p27_231")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(13,14),group2=c(15,16),name1="p27_231_DD", name2="p27_1833")
tamoxifen <- dba.contrast(tamoxifen, group1 = c(5,6),group2=c(15,16),name1="p27_231", name2="p27_1833")
tamoxifen <- dba.analyze(tamoxifen)
save(tamoxifen,file=file.path(output.file.dir,"Danny_Dba.RData"))
tamoxifen
# dba.report(tamoxifen,contrast = 1)
# dba.report(tamoxifen,contrast = 2)
# dba.report(tamoxifen,contrast = 3)
# dba.report(tamoxifen,contrast = 4)
# dba.report(tamoxifen,contrast = 5)
# dba.report(tamoxifen,contrast = 6)
}
#generateBedFromDba(tamoxifen,"~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed")
generateBedFromDba <- function(tamoxifen,output.file.dir){
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
n <- length(tamoxifen$contrasts)
lapply(1:n,function(u,tamoxifen){
temp <- as.data.frame(dba.report(tamoxifen,contrast = u,th=1))
file.name <- paste(colnames(as.data.frame(dba.report(tamoxifen,contrast = u,th=1)))[c(7,8)],collapse = "-")
write.table(temp, row.names=F, col.names = F, quote =F,sep="\t", file=file.path(output.file.dir,paste0(file.name,".bed")))
},tamoxifen)
}
#' dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed"
#' input.file.pattern=".bed"
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker2(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnntationUsingChipSeeker2 <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=5000,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
re<-ParserReadFiles(dir.name,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2 <- re.bed
# if(length(dir(dir.name,pattern="peaks.bed"))!=0)
# {
# re.peaks.only.bed.2<-FL(re.bed,'peaks')
# cat("peaks\n")
# print(re.peaks.only.bed.2)
# }
#
# if(length(dir(dir.name,pattern="summits.bed"))!=0){
# re.summits.only.bed<-FL(re.bed,'summits')
# cat("summits\n")
# print(re.summits.only.bed)
# }
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,"Annotation",APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
peaks.anno.list <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
#select.index <- which(peakAnno$distanceToTSS<20,000 && peakAnno$distanceToTSS >= -20,000)
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
x_name=names(re.peaks.only.bed.2)[u]
cat(x_name,"\n")
png(file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_pie.png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
#print(paste0(peaks[,c(2,3)]))
group1 <- strsplit(tools::file_path_sans_ext(x_name),"-")[[1]][1]
group2 <- strsplit(tools::file_path_sans_ext(x_name),"-")[[1]][2]
colnames(peaks.anno)[5:13]=c("starnd","width_DiffDind_based","strand","Conc", group1,group2,"Fold","p.value","FDR")
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_only_mapped_peaks.xls")),
row.names = FALSE,quote=FALSE,sep="\t")
# unmapped.peaks<-peaks[-which(paste0(peaks[,2],"_",peaks[,3]) %in% paste0(peaks.anno[,2],"_",peaks.anno[,3])),]
#
# cat(dim(peaks)[1]," ",dim(peaks.anno)[1]," ",dim(unmapped.peaks)[1],"\n")
#
#
# if(dim(unmapped.peaks)[1]!=0){
#
# colnames(unmapped.peaks)=colnames(peaks.anno)[1:6]
#
# unmapped.peaks.3<-smartbind(peaks.anno,unmapped.peaks)
#
# unmapped.peaks.4<-unmapped.peaks.3[order(unmapped.peaks.3[,1],unmapped.peaks.3[,2]),]
#
# write.table(unmapped.peaks.4,file=file.path(temp3,paste0(x_name,"_",d,APpath,"_around_tss_annotation_4_all_peaks.xls")),row.names = FALSE,quote=FALSE,sep="\t")
# }
#
peaks.anno
},re.peaks.only.bed.2,d)
}
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/CombineChipSeqWithRNASeq"
#' rna.seq.input.file="/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx"
#'
#' integrateChipSeqWithRNASeq(out.dir.name,rna.seq.input.file)
#'
integrateChipSeqWithRNASeq <- function(output.file.dir,rna.seq.input.file) {
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
gene.231dd.231.1833.1833shp27 <- read.xlsx(rna.seq.input.file,sheetIndex = 1,header = FALSE)
colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
data.231DD.231 <- gene.231dd.231.1833.1833shp27[,c(1,2,3)]
data.1833.231 <- gene.231dd.231.1833.1833shp27[,c(1,4,5)]
data.1833shp27.1833 <- gene.231dd.231.1833.1833shp27[c(1,6,7)]
cJun.231DD.231.ChipSeq.RNASeq <- merge(res.promoter[[2]],data.231DD.231,by="SYMBOL")
p27.231DD.231.ChipSeq.RNASeq <- merge(res.promoter[[5]],data.231DD.231,by="SYMBOL")
cJun.231.1833.ChipSeq.RNASeq <- merge(res.promoter[[3]],data.1833.231,by="SYMBOL")
p27.231.1833.ChipSeq.RNASeq <- merge(res.promoter[[6]],data.1833.231,by="SYMBOL")
write.table(cJun.231DD.231.ChipSeq.RNASeq,file=file.path(output.file.dir,"cJun_231DD_231_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(p27.231DD.231.ChipSeq.RNASeq,file=file.path(output.file.dir,"p27_231DD_231_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(cJun.231.1833.ChipSeq.RNASeq,file=file.path(output.file.dir,"cJun_231_1833_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
write.table(p27.231.1833.ChipSeq.RNASeq,file=file.path(output.file.dir,"p27_231_1833_DBA_DGE.xls"),
row.names = FALSE,quote=FALSE,sep="\t")
}
addTestFunction4HeatmapChipSeq <- function(GenomicRanges, rtracklayer, IRanges, dataDirectory, chipseq, VennDiagram) {
library(GenomicRanges)
library(rtracklayer)
library(IRanges)
input = import.bed(file.path(dataDirectory, 'ES_input_filtered_ucsc_chr6.bed'),
asRangedData=FALSE)
rep1 = import.bed("/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/2017-03-02-01_S5_R1-MDA-MB-231-DD-1-cJun_hs_1.00e-05_macs142_peaks.bed",genome = "hg19")
rep2 = import.bed("/Volumes/Bioinformatics$/2017/DannyNewData/PeakCall/2017-03-02-02_S2_R1-MDA-MB-231-DD-2-cJun_hs_1.00e-05_macs142_peaks.bed",genome = "hg19")
library(chipseq)
estimate.mean.fraglen(rep1)
prepareChIPseq = function(reads){
frag.len = median( estimate.mean.fraglen(reads) )
cat( paste0( 'Median fragment size for this library is ', round(frag.len)))
reads.extended = resize(reads, width = frag.len)
return( trim(reads.extended) )
}
rep1 = prepareChIPseq(rep1)
ovlp = findOverlaps(rep1,rep2)
ovlp = as.data.frame(ovlp)
ov = min( length(unique(ovlp$queryHits)), length(unique(ovlp$subjectHits)) )
library(VennDiagram)
draw.pairwise.venn(
area1=length(rep1),
area2=length(rep2),
cross.area=ov,
category=c("rep1", "rep2"),
fill=c("steelblue", "blue3"),
cat.cex=0.7)
}
#input.region.bed.dir = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
#plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin","hg19_gene.bed",)
#
plotHeatMapUsedeepTools <- function(input.sample.file,input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
bw.file.sample.label <- mapBw3Sample(input.sample.file,input.bw.file.dir)
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
bed.file.list <- list.files(
input.region.bed.dir,
pattern = "*.bed$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
bw.file.list <- as.character(bw.file.sample.label$file.bw)
samplesLabel <- as.character(bw.file.sample.label$sampleLabel)
print(bed.file.list)
if(select.region.bed!=""){
input.beds = bed.file.list[-grep(paste(select.region.bed,collapse = "|"), bed.file.list)]
input.beds = paste(input.beds,collapse = " ")
}else{
input.beds = paste(bed.file.list,collapse = " ")
}
print(input.beds)
cmd = "computeMatrix reference-point --referencePoint TSS -b 4000 -a 4000 -R"
input.beds=input.beds
input.bw.file=paste(bw.file.list,collapse = " ")
input.samplesLabel=paste(samplesLabel,collapse = " ")
outFileNameMatrix=file.path(output.file.dir,"matrix1_cJun_p27_TSS.gz")
outFileSortedRegions=file.path(output.file.dir,"regions_cJun_p27_genes.bed")
outHeatMapFile=file.path(output.file.dir,"heatmap_cJun_p27.png")
cmd1=paste(cmd,input.beds,"-S",input.bw.file,"--skipZero","-o",outFileNameMatrix,"--outFileSortedRegions",outFileSortedRegions,"--samplesLabel",input.samplesLabel,collapse = " ")
cmd2=paste("plotHeatmap -m",outFileNameMatrix,"-out",outHeatMapFile,collapse = " ")
#
# testFiles/genes.bed -S testFiles/log2ratio_H3K4Me3_chr19.bw --skipZeros
# -o matrix1_H3K4me3_l2r_TSS.gz
# --outFileSortedRegions regions1_H3K4me3_l2r_genes.bed
#
# deepTools2.0/bin/computeMatrix scale-regions \
# -R genes_chr19_firstHalf.bed genes_chr19_secondHalf.bed \ # separate multiple files with spaces
# -S testFiles/log2ratio_*.bw \ or use the wild card approach
# -b 3000 -a 3000 \
# --regionBodyLength 5000 \
# --skipZeros -o matrix2_multipleBW_l2r_twoGroups_scaled.gz \
# --outFileNameMatrix matrix2_multipleBW_l2r_twoGroups_scaled.tab \
# --outFileSortedRegions regions2_multipleBW_l2r_twoGroups_genes.bed
#
# computeMatrix reference-point \ # choose the mode
# --referencePoint TSS \ # alternatives: TES, center
# -b 3000 -a 10000 \ # define the region you are interested in
# -R testFiles/genes.bed \
# -S testFiles/log2ratio_H3K4Me3_chr19.bw \
# --skipZeros \
# -o matrix1_H3K4me3_l2r_TSS.gz \ # to be used with plotHeatmap and plotProfile
# --outFileSortedRegions regions1_H3K4me3_l2r_genes.bed
#cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd1=paste(cmd.java.2,cmd1,sep=";")
system(cmd1)
#print(cmd2)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin/annotation","hg19_gene.bed","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4plotHeatMapUsedeepTools <- function(input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "HeapMapPlot"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::plotHeatMapUsedeepTools('
Rinput <- paste0('\\"',input.bw.file.dir,'\\",',
'\\"',input.region.bed.dir,'\\",',
'\\"',select.region.bed,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
}
# Welcome to Rattler, *please* read these important system notes:
#
# --> Rattler is currently running the SLURM resource manager to
# schedule all compute resources. Example SLURM job scripts are
# available on the system at /cm/shared/docs/slurm/current/job_scripts
#
# To run an interactive shell, issue:
# srun -p hipri -t 0:30:00 -n 36 -N 1 --pty /bin/bash -l
#
# To submit a batch job, issue: sbatch job_script
# To show all queued jobs, issue: squeue
# To kill a queued job, issue: scancel <jobId>
#
# See "man slurm" for more detailed information.
#
# --> Rattler has 3 main queues with the following timelimits:
# * hipri (2 days)
# * iq (1 day)
# * requestq (infinite) - Requires a valid slurm reservation to use
#
# --> To see which software packages are available issue: "module avail"
#
# Use the following commands to adjust your environment:
# 'module add <module>' - adds a module to your environment for this session
# 'module initadd <module>' - configure module to be loaded at every login
#
#plotHeatMapUsedeepTools("/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap/matrix1_cJun_p27_TSS.gz","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")
#
usePlotHeatmap <- function(input.matrix.file,output.file.dir){
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
outHeatMapFile=file.path(output.file.dir,"heatmap_cJun_p27.png")
cmd2=paste("plotHeatmap -m",input.matrix.file,"-out",outHeatMapFile,collapse = " ")
system(cmd2)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4usePlotHeatmap("/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap/matrix1_cJun_p27_TSS.gz","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4usePlotHeatmap <- function(input.matrix.file,output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "plotHeatmap"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::usePlotHeatmap('
Rinput <- paste0('\\"',input.matrix.file,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
system(cmd.gff)
}
# input.bam.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed/Annotation"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
# generateBed4HeatMap(input.bam.file.dir,out.dir.name)
#
generateBed4HeatMap <- function(input.bam.file.dir,out.dir.name) {
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.list <- list.files(
input.bam.file.dir,
pattern = "*xls$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
file.table <- lapply(file.list,function(u){
y <- basename(u)
pos <- regexpr("\\_", y)
pos <- pos - 1
y <- substr(y, 1, pos)
x <-fread(u)
x<- cbind(x,rep(y,dim(x)[1]))
colnames(x)[dim(x)[2]]="sample_name"
x
})
# library(dplyr)
# x <- data_frame(i = c("a","b","c"), j = 1:3)
# y <- data_frame(i = c("b","c","d"), k = 4:6)
# z <- data_frame(i = c("c","d","a"), l = 7:9)
common.gene.peak <- file.table %>%
Reduce(function(dtf1,dtf2) inner_join(dtf1,dtf2,by="SYMBOL"),.)
x <- data.table::subset(common.gene.peak,select=c("geneChr.x","geneStart.x","geneEnd.x","geneStrand.x","SYMBOL"))
data.table::setkey(x,NULL)
xx <- unique(x)
xxx<-xx[complete.cases(xx),]
# gene.231dd.231.1833.1833shp27 <- read.xlsx(
# "/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx",
# sheetIndex = 1,header = FALSE)
# colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
#save(gene.231dd.231.1833.1833shp27,file="./data/gene.RData")
gene.45 <- merge(gene.231dd.231.1833.1833shp27,xxx,by="SYMBOL",sort = FALSE)
gene.45.1 <- gene.45[,c(8,9,10,11,1)]
colnames(gene.45.1)=c("chr","start","end","strand","SYMBOL")
gene.45.1[which(gene.45.1$strand==1),]$strand="+"
gene.45.1[which(gene.45.1$strand==2),]$strand="-"
gene.45.1.sorted.by.chr <- gene.45.1[order(gene.45.1$chr),]
gene.45.1$chr<-paste0("chr",gene.45.1$chr)
gene.45.1.sorted.by.chr$chr<-paste0("chr",gene.45.1.sorted.by.chr$chr)
write.table(gene.45.1,file=file.path(out.dir.name,"gene_200.bed"),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
write.table(gene.45.1.sorted.by.chr,file=file.path(out.dir.name,"gene_200_sorted.bed"),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("~/SampleID_INFO_ChIP_new_Danny.csv","~/BamCompare","~/ChipSeqBed",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmapAddlabel")'
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("~/SampleID_INFO_ChIP_new_Danny.csv","~/BamCoverage","~/ChipSeqBed_p27_cJun",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap4p27_cJun")'
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4plotHeatMapUsedeepTools("/nethome/axy148/R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","~/BamCompareZhao","~/cJun_gene_bed",select.region.bed=NULL,"/scratch/projects/bbc/aiminy_project/zhao_ChipSeq_heatmap4cJun")'
bashJob4plotHeatMapUsedeepTools <- function(input.sample.file,input.bw.file.dir,input.region.bed.dir,select.region.bed,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "generateMatrix"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::plotHeatMapUsedeepTools('
Rinput <- paste0('\\"',input.sample.file,'\\",',
'\\"',input.bw.file.dir,'\\",',
'\\"',input.region.bed.dir,'\\",',
'\\"',select.region.bed,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
job.name <- "plotHeatmap"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::usePlotHeatmap('
input.matrix.file <- file.path(output.file.dir,"matrix1_cJun_p27_TSS.gz")
Rinput <- paste0('\\"',input.matrix.file,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name="generateMatrix")
system(cmd.gff)
}
#input.bw.dir="~/BamCompare"
#input.sample.file="~/SampleID_INFO_ChIP_new_Danny.csv"
#
#mapBw3Sample(input.sample.file,input.bw.dir)
#
mapBw3Sample <- function(input.sample.file,input.bw.dir) {
file.1 <- list.files(input.bw.dir,pattern=".bw$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p1 <- regexpr("\\_", x);p2 <- regexpr("\\.", x);xx <- substr(x,p1+1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bw")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
makeHeatMapByme <- function() {
compute.matrix.file="/Users/axy148/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Danny_ChipSeq_heatmapBasedSortedBed/matrix1_cJun_p27_TSS.gz"
table = read.table(gzfile(compute.matrix.file), skip=1)
matrix = table[7:dim(table)[2]]
image(1:dim(matrix)[2], 1:dim(matrix)[1], t(as.matrix(matrix)), axes=FALSE, xlab='sample', ylab='gene')
axis(2, at=1:length(table$V1), labels=table$V4, las = 1)
}
# input.bam.file.dir="/Volumes/Bioinformatics$/2017/DannyNewData/BindDiff/common_peaks_bed/Annotation"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name)
#
# input.bam.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed/p27"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name,"p27")
#
# input.bam.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed/cJun"
# out.dir.name = "~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
# gene.full <- generateBed4HeatMap2(input.bam.file.dir,out.dir.name,"cJun")
#
generateBed4HeatMap2 <- function(input.bam.file.dir,out.dir.name,Ab) {
out.dir.name <-file.path(out.dir.name,paste0(Ab,"_gene_bed"))
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.list <- list.files(
input.bam.file.dir,
pattern = "*xls$",
all.files = TRUE,
full.names = TRUE,
recursive = TRUE,
include.dirs = TRUE)
file.table <- lapply(file.list,function(u){
y <- basename(u)
pos <- regexpr("\\_", y)
pos <- pos - 1
y <- substr(y, 1, pos)
x <-fread(u)
x<- cbind(x,rep(y,dim(x)[1]))
colnames(x)[dim(x)[2]]="sample_name"
x
})
# library(dplyr)
# x <- data_frame(i = c("a","b","c"), j = 1:3)
# y <- data_frame(i = c("b","c","d"), k = 4:6)
# z <- data_frame(i = c("c","d","a"), l = 7:9)
common.gene.peak <- file.table %>%
Reduce(function(dtf1,dtf2) full_join(dtf1,dtf2,by="SYMBOL"),.)
x <- subset(common.gene.peak,select=c("geneChr.x","geneStart.x","geneEnd.x","geneStrand.x","SYMBOL"))
setkey(x,NULL)
xx <- unique(x)
xxx<-xx[complete.cases(xx),]
# gene.231dd.231.1833.1833shp27 <- read.xlsx(
# "/Volumes/Bioinformatics$/2017/DannyNewData/AnnotationNew4DBA/table_S3_headerless.xlsx",
# sheetIndex = 1,header = FALSE)
# colnames(gene.231dd.231.1833.1833shp27)=c("SYMBOL","FC_231DD_231","p_231DD_231","FC_1833_231","p_1833_231","FC_1833shp27_1833","p_1833shp27_1833")
#save(gene.231dd.231.1833.1833shp27,file="./data/gene.RData")
#gene.45 <- merge(gene.231dd.231.1833.1833shp27,xxx,by="SYMBOL",sort = FALSE)
gene.45 <- xxx
gene.45.1 <- gene.45
colnames(gene.45.1)=c("chr","start","end","strand","SYMBOL")
gene.45.1[which(gene.45.1$strand==1),]$strand="+"
gene.45.1[which(gene.45.1$strand==2),]$strand="-"
gene.45.1.sorted.by.chr <- gene.45.1[order(gene.45.1$chr),]
gene.45.1$chr<-paste0("chr",gene.45.1$chr)
gene.45.1.sorted.by.chr$chr<-paste0("chr",gene.45.1.sorted.by.chr$chr)
write.table(gene.45.1,file=file.path(out.dir.name,paste0("gene_",Ab,".bed")),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
write.table(gene.45.1.sorted.by.chr,file=file.path(out.dir.name,paste0("gene_",Ab,"_sorted.bed")),row.names = FALSE,col.names = FALSE,quote=FALSE,sep="\t")
# > length(unique(file.table[[1]]$SYMBOL))
# [1] 417
# > length(unique(file.table[[2]]$SYMBOL))
# [1] 222
# > length(unique(file.table[[3]]$SYMBOL))
# [1] 458
#
#
# unique(c(unique(file.table[[1]]$SYMBOL),unique(file.table[[2]]$SYMBOL),unique(file.table[[3]]$SYMBOL)))
gene.45.1.sorted.by.chr
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "BamCoverage" -o %J.BamCoverage.log -e %J.BamCoverage.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCoverage("/scratch/projects/bbc/aiminy_project/DannyNewData2/SampleID_INFO_ChIP_new_Danny.csv","/nethome/axy148/_visualization/sorted_bam_files_2.txt","~/BamCoverage")'
useBamCoverage <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
#bamCoverage -b reads.bam -o coverage.bw
#bamCoverage --bam a.bam -o a.SeqDepthNorm.bw \
#--binSize 10
#--normalizeTo1x 2150570000
#--ignoreForNormalization chrX
#--extendReads
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX"
cmd3 <- "-o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("To1x_",basename(as.character(xx)),"_vs_","no_input",".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
#' output.file.dir="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/heatmap/Peak2Bed"
#' #mcf7=mcf2
#' re<-getTargetGene4Ab(mcf2,"yes","p27",output.file.dir)
#' re<-getTargetGene4Ab(mcf2,"yes","cJun",output.file.dir)
#'
getTargetGene4Ab<-function(mcf7,Mergereplicates=c("yes","no"),Ab,output.file.dir){
output.file.dir = file.path(output.file.dir,Ab)
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir,recursive = TRUE)
}
temp<-mcf7
sampID.v<-colnames(temp$class)
sampID.v.2<-unlist(lapply(1:length(sampID.v),function(u,sampID.v){
x=sampID.v[u]
y=x
y
},sampID.v))
colnames(temp$class)<-sampID.v.2
temp$class[1,]<-sampID.v.2
#temp<-dba(temp,mask =c(1,2,13,14,11,12,15,16))
#Merge the replicates of each set
if(Mergereplicates=="yes"){
temp2<-dba.peakset(temp,consensus = -DBA_REPLICATE)
#temp22<-dba(temp2,mask =c(9,16,17:23))
print(temp2)
##need to set the flexiable number identified from data sets
t <- temp2
temp22<-dba(t,mask =which(t$class[which(rownames(t$class)=="Replicate"),]=="1-2"))
}else{
temp22<-temp
}
print(temp22)
#temp22 <- re
Tissue1<-temp22$class[2,]
Tissue2<-unique(temp22$class[2,])
TF<-unique(temp22$class[3,])
TF.n<-length(TF)
for(i in 1:length(Tissue2)){
#for(j in 1:length(TF)){
p <- which(temp22$class[2,]==Tissue2[i]&temp22$class[3,]==Ab)
cat(Tissue2[i]," ",Ab," ",p,"\n")
if(length(p)!=0){
#as.data.frame(dba.peakset(temp22,peaks=1,bRetrieve=TRUE))
dba.peakset(temp22,peaks=as.integer(p),bRetrieve=TRUE, writeFile = file.path(output.file.dir,paste0(Tissue2[i],"_",Ab,".bed")))
}
}
AnntationUsingChipSeeker3(output.file.dir,".bed",output.file.dir
,txdb="hg19",DD=5000)
# AnntationUsingChipSeeker2
# temp3=file.path(output.file.dir,"Venn")
#
# if(!dir.exists(temp3))
# {
# dir.create(temp3)
# }
#
# for(i in 1:length(Tissue2)){
# po<-which(Tissue1 %in% Tissue2[i])
#
# print(po)
#
# if(length(po)==2)
# {
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po,main=Tissue2[i])
# dev.off()
#
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po1],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po1,main=Tissue2[i])
# dev.off()
#
# png(file.path(temp3,paste0(paste0(colnames(temp22$class)[po2],collapse = "-vs-"),".png")))
# dba.plotVenn(temp22,mask=po2,main=Tissue2[i])
# dev.off()
#
# }else
# {
# cat(paste0("For ",Tissue2[i],": Only one peak profile for ",TF.n," TFs\n"))
# }
# }
#
# p.common<-lapply(1:length(Tissue2),function(u,Tissue1,Tissue2,temp22){
#
# po<-which(Tissue1 %in% Tissue2[u])
#
# if(length(po)==2)
# {
# common.peaks<-dba.overlap(temp22,mask=po)
# y<-common.peaks$inAll
# }else if(length(po)==4){
# po1<-po[c(1,3)]
# po2<-po[c(2,4)]
#
# common.peaks.1<-dba.overlap(temp22,mask=po1)
# y1<-common.peaks.1$inAll
#
# common.peaks.2<-dba.overlap(temp22,mask=po2)
# y2<-common.peaks.2$inAll
#
# y<-list(y1=y1,y2=y2)
#
# names(y)[1]<-paste0(colnames(temp22$class)[po1],collapse = "-vs-")
# names(y)[2]<-paste0(colnames(temp22$class)[po2],collapse = "-vs-")
#
# }else{
# y<-NULL}
# y
# },Tissue1,Tissue2,temp22)
#
# names(p.common)<-Tissue2
#
# p.common<-unlist(p.common,recursive = F)
#
# p.common<-p.common[lapply(p.common,length) > 0]
#
# if(!dir.exists(file.path(output.file.dir,"common_peaks_bed")))
# {
# dir.create(file.path(output.file.dir,"common_peaks_bed"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","ucsc"))
# dir.create(file.path(output.file.dir,"common_peaks_bed","igv"))
# }
#
# #output common peaks to bed files
#
# lapply(1:length(p.common),function(u,p.common,output.file.dir){
# x=p.common[[u]]
#
# x_name=names(p.common)[u]
#
# df <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=strand(x))
#
# #assign strand
# df.str <- data.frame(seqnames=seqnames(x),
# #starts=start(x)-1,
# starts=start(x),
# ends=end(x),
# names=c(rep(".", length(x))),
# scores=elementMetadata(x)[,1],
# strands=c(rep(".", length(x))))
#
# df.str.1<-df.str[-grep("random",df.str$seqnames),]
#
# df.str.2<-df.str.1
#
# df.str.3<-df.str.2[-grep("chrUn",df.str.2$seqnames),]
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_cp_with_header.bed")),
# col.names=TRUE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","ucsc",paste0(x_name,"_4_ucsc.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed",paste0(x_name,"_common_peaks.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# write.table(df,file=file.path(output.file.dir,"common_peaks_bed","igv",paste0(x_name,"_4_igv.bed")),
# col.names=FALSE,row.names = FALSE,quote=FALSE,sep="\t")
#
# },p.common,output.file.dir)
#
# AnntationUsingChipSeeker(file.path(output.file.dir,"common_peaks_bed","igv"),"bed",file.path(output.file.dir,"common_peaks_bed")
# ,txdb="hg19",DD=5000,distanceToTSS_cutoff=10000)
#
# return(p.common)
}
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("~/SampleID_INFO_ChIP_new_Danny.csv","~/sort_bam","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_data_QC")'
#
#R/lib64/R/library/ChipSeq/extdata/zhao_data.csv
#
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","~/sort_bam","/scratch/projects/bbc/aiminy_project/zhao_ChipSeq_data_QC")'
#
#bsub -P bbc -J "RunSppR" -o %J.RunSppR.log -e %J.RunSppR.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useRunSppR("~/SampleID_INFO_ChIP_new_Danny.csv","/projects/scratch/bbc/Project/Danny_chip2/BigWig","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_data_QC_2")'
#
#
#
useRunSppR <- function(input.sample.file,input.bam.file.dir,output.file.dir){
#bam.file.sample.label <- mapBam2Sample2(input.sample.file,input.bam.file.dir)
bam.file.sample.label <- mapBam2Sample3(input.sample.file,input.bam.file.dir)
if(!dir.exists(output.file.dir)){dir.create(output.file.dir,recursive = TRUE)}
#/projects/ctsi/bbc/aimin/annotation/hg19_gene.bed
bam.file.list <- as.character(bam.file.sample.label$file.bam)
samplesLabel <- as.character(bam.file.sample.label$sampleLabel)
lapply(1:length(bam.file.list),function(u,bam.file.list,samplesLabel,output.file.dir){
cmd = paste("Rscript ~/phantompeakqualtools/run_spp.R",
paste0("-c=",bam.file.list[u]),
paste0("-savp=",file.path(output.file.dir,paste0(samplesLabel[u],"_QC.pdf"))),
paste0("-out=",file.path(output.file.dir,paste0(samplesLabel[u],"_QC_metrix.txt"))),sep=" ")
system(cmd)
},bam.file.list,samplesLabel,output.file.dir)
}
#R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::submitJob4plotHeatMapUsedeepTools("~/BamCompare","/projects/ctsi/bbc/aimin/annotation","hg19_gene.bed","/scratch/projects/bbc/aiminy_project/Danny_ChipSeq_heatmap")'
submitJob4useRunSppR <- function(input.sample.file,input.bam.file.dir,output.file.dir){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
job.name <- "QC"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::useRunSppR('
Rinput <- paste0('\\"',input.sample.file,'\\",',
'\\"',input.bam.file.dir,'\\",',
'\\"',output.file.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
cmd.gff <- DoGs:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
system(cmd.gff)
}
#input.bw.dir="~/BamCompare"
#input.sample.file="~/SampleID_INFO_ChIP_new_Danny.csv"
#
#mapBw3Sample(input.sample.file,input.bw.dir)
#
mapBam2Sample <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p1 <- regexpr("\\_", x);p2 <- regexpr("\\.", x);xx <- substr(x,p1+1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
mapBam2Sample2 <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);p2 <- regexpr("\\.", x);xx <- substr(x,1,p2-1)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
mapBam2Sample3 <- function(input.sample.file,input.bam.dir) {
file.1 <- list.files(input.bam.dir,pattern=".bam$",all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
sample.file <- fread(input.sample.file)
file.2<-cbind(unlist(lapply(file.1,function(u){x<-basename(u);xx <- tools::file_path_sans_ext(x)})),file.1)
colnames(file.2)=c("ID","file.bam")
file.3 <- merge(file.2,sample.file,by="ID",sort=F)
sampleLabel= paste(gsub(" ", "-", file.3$Type_Cell), file.3$Type_TF, sep = "-")
sampleLabel=gsub("MDA-MB-","",sampleLabel)
file.4 <- cbind(file.3,sampleLabel)
file.5 <- file.4[,c(2,6)]
file.5
}
#' dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/Bba2Bed"
#' input.file.pattern=".bed"
#' out.dir.name="~/Dropbox (BBSR)/BBSR Team Folder/Aimin_Yan/ChipSeq/AnnotationNew4DBA"
#' txdb="hg19"
#' DD=5000
#'
#' AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000, AP=c("Promoter","Intron"))
#'
#' res.promoter <- AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Promoter"))
#'
#' AnntationUsingChipSeeker3(dir.name,input.file.pattern,out.dir.name,txdb=txdb,DD,distanceToTSS_cutoff=5000,AP=c("Intron"))
#'
AnotationUsingChipSeeker3 <- function(dir.name,input.file.pattern,out.dir.name,txdb=c("hg19","hg38"),DD,distanceToTSS_cutoff=NULL,assignGenomicAnnotation=TRUE,AP=c("Promoter", "5UTR", "3UTR", "Exon", "Intron","Downstream", "Intergenic")) {
re<-ParserReadFiles(dir.name,input.file.pattern)
re.bed<-re$input
re.peaks.only.bed.2 <- re.bed
txdb<-match.arg(txdb)
switch (txdb,
hg38 = {
cat("Use hg38\n")
txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
},
{
cat("Use hg19\n")
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
}
)
APpath <- paste(AP,collapse = "_")
temp3=file.path(out.dir.name,APpath)
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
d=DD
peaks.anno.list <- lapply(1:length(re.peaks.only.bed.2),function(u,re.peaks.only.bed.2,d){
peaks=readPeakFile(re.peaks.only.bed.2[[u]],as="data.frame")
print(head(peaks))
if(dim(peaks)[1]>0){
peakAnno <- annotatePeak(re.peaks.only.bed.2[[u]], tssRegion=c(-d, d),
TxDb=txdb,assignGenomicAnnotation=assignGenomicAnnotation,genomicAnnotationPriority=AP,annoDb="org.Hs.eg.db")
dropAnnoM <- function (csAnno, distanceToTSS_cutoff)
{
idx <- which(abs(mcols(csAnno@anno)[["distanceToTSS"]]) <
distanceToTSS_cutoff)
csAnno@anno <- csAnno@anno[idx]
csAnno@peakNum <- length(idx)
if (csAnno@hasGenomicAnnotation) {
csAnno@annoStat <- ChIPseeker:::getGenomicAnnoStat(csAnno@anno)
csAnno@detailGenomicAnnotation = csAnno@detailGenomicAnnotation[idx,
]
}
csAnno
}
if(!is.null(distanceToTSS_cutoff)){
peakAnno <- dropAnnoM(peakAnno,distanceToTSS_cutoff = distanceToTSS_cutoff)
}else
{
peakAnno <- peakAnno
}
x_name=names(re.peaks.only.bed.2)[u]
x_name=tools::file_path_sans_ext(x_name)
x_name=tools::file_path_sans_ext(x_name)
cat(x_name,"\n")
png(file.path(temp3,paste0(x_name,"_",d,".png")))
plotAnnoPie(peakAnno)
dev.off()
peaks.anno=as.data.frame(peakAnno)
print(head(peaks.anno))
print(colnames(peaks.anno))
write.table(peaks.anno,file=file.path(temp3,paste0(x_name,"_",d,".xls")),
row.names = FALSE,quote=FALSE,sep="\t")
peaks.anno
}
},re.peaks.only.bed.2,d)
}
# R -e 'library(PathwaySplice);library(ChipSeq);ChipSeq:::bashJob4generateBed4HeatMap2("~/cJun","~/","cJun")'
bashJob4generateBed4HeatMap2 <- function(input.bam.file.dir,out.dir.name,Ab){
#Sys.setenv(JAVA_HOME='/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server')
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
job.name <- "generateGeneBed"
Rfun1 <- 'library(ChipSeq);re <- ChipSeq:::generateBed4HeatMap2('
Rinput <- paste0('\\"',input.bam.file.dir,'\\",',
'\\"',out.dir.name,'\\",',
'\\"',Ab,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
#cmd.java.1="module load java/1.8.0_60"
#cmd.java.1="R CMD javareconf -e"
# cmd.java.2="export LD_LIBRARY_PATH=/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre/lib/amd64/server:$LD_LIBRARY_PATH"
#cmd.java ='export JAVA_HOME="/usr/lib/jvm/java-1.7.0-openjdk-1.7.0.45.x86_64/jre"'
cmd.gff <- PathwaySplice:::createBsubJobArrayRfun(Rfun,job.name,wait.job.name=NULL)
#cmd2=paste(cmd.java.2,cmd.gff,sep=";")
system(cmd.gff)
}
# /deepTools-1.5/bin/bamCompare --bamfile1 ChIP.bam --bamfile2 Input.bam \
# --binSize 25 --fragmentLength 200 --missingDataAsZero no \
# --ratio log2 --scaleFactorsMethod SES -o log2ratio_ChIP_vs_Input.bw
#' bsub -P bbc -J "bamCompare" -o %J.bamCompare.log -e %J.Compare.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::useBamCompare2("/nethome/axy148/R/lib64/R/library/ChipSeq/extdata/zhao_data.csv","/nethome/axy148/_visualization/sort_bam.txt","~/BamCompareZhao")'
useBamCompare2 <- function(input.sample.file,input.bam.file,output.dir)
{
#sampleF <- read.table(input.sample.file,header = FALSE)
#colnames(sampleF) <- c("ID","SampleName")
#bam <- prepareBamFile(input.bam.file,file.pattern)
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1",xx,"--bamfile2",x.input,sep=" ")
cmd2 <- "--binSize 25"
cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(x.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
system("module unload python/2.7.3")
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::ngs()'
ngs <- function(){
system("ngs.plot.r -G hg19 -R tss -C ~/zhao_config.txt -O zhao.tss -L 4000")
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::ngs2()'
ngs2 <- function(config.file,distance.around.tss,output.file.dir){
if (!dir.exists(output.file.dir))
{
dir.create(output.file.dir, recursive = TRUE)
}
cmd=paste("ngs.plot.r -G hg19 -R tss -C",config.file,"-O",output.file.dir,"-L",distance.around.tss,"-CO white:white:red", collapse = " ")
system(cmd)
}
doAlignment<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
filterSam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd2='samtools view -Sh /home/someusername/myFastqFile.sam | grep -e "^@" -e "XM:i:[012][^0-9]" | grep -v "XS:i:" > /home/someusername/myFastqFile.sam.filtered.sam'
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
convertSam2Bam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd3="samtools view -S -b /home/someusername/myFastqFile.sam.filtered.sam > /home/someusername/myFastqFile.sam.filtered.sam.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
sortBam<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd4="samtools sort /home/someusername/myFastqFile.sam.filtered.sam.bam /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
removePCRduplicates<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd5="samtools rmdup -s /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
indexBAM<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd6="samtools index /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
bam2Bed<-function(input.fastq.dir,bowtie2Indeces,output.dir){
re <- parserreadfiles(input.fastq.dir, "fastq")
res <- re$input
index <- system('echo $LSB_JOBINDEX',intern = TRUE)
cmd7="bedtools bamtobed -i /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam > /home/someusername/myFastqFile.sam.filtered.sam.bam.sorted.bam.nodup.bam.bed"
cmd0="bowtie2 -x"
"/home/someusername/bowtie2Indeces/somegenome -U /home/someusername/myFastqFile.fastq -S /home/someusername/myFastqFile.sam"
u <- as.integer(index)
path_name = dirname(res[[u]])
file_name = file_path_sans_ext(basename(res[[u]]))
cmd1 <- paste(cmd0,bowtie2Indeces,"-U",res[[u]],"-S",file.path(output.dir,paste0(file_name,".sam")), sep = " ")
cmd2 <- cmd1
system(cmd2)
cat(cmd2,"\n")
}
getBamReady<-function(input.fastq.dir,bowtie2Indecesinput,sample.num,output.bam.dir){
Rfun1 <- 'library(ChipSeq);library(DoGs);re <- ChipSeq:::doAlignment('
Rinput <- paste0('\\"',input.fastq.dir,'\\",',
'\\"',bowtie2Indecesinput,'\\",',
'\\"',output.dir,'\\"')
Rfun2 <- ')'
Rfun <-paste0(Rfun1,Rinput,Rfun2)
alignment <- createBsubJobArrayRfun(Rfun,paste0("alignment[1-",sample.num,"]",NULL))
system(alignment)
}
getEffectOfRmDup<-function(){
not.rm <- read.table("~/Dropbox (BBSR)/Aimin_project/Research/ChipSeq/Danny_ChipSeq_data_QC/qc_16.txt")
rm <- read.table("~/Dropbox (BBSR)/Aimin_project/Research/ChipSeq/Danny_ChipSeq_data_QC_2/16_samples_QC.txt")
res <- as.data.table(rbind(cbind(rep("not_rm",length(not.rm[,11])),not.rm[,11]),cbind(rep("rm",length(rm[,11])),rm[,11])))
boxplot(as.numeric(res$V2)~res$V1)
}
#' sbatch -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::runIDR()'
#' sbatch -p hipri test-job.sh
#' sbatch -p hipri test-job.sh
#' squeue -u steven.miamiuniv
runIDR <- function()
{
cmd="idr --samples /home/steven.miamiuniv/Softwares/idr-2.0.2/tests/data/peak1 /home/steven.miamiuniv/Softwares/idr-2.0.2/tests/data/peak2 --plot testIDR"
system(cmd)
}
# bsub -P bbc -J "zhaoCJun" -o %J.zhaoCJun.log -e %J.zhaoCJun.err -W 72:00 -n 32 -q parallel -R 'rusage[mem= 16000 ] span[ptile= 16 ]' -u aimin.yan@med.miami.edu R -e 'library(ChipSeq);re <- ChipSeq:::peakCallAndAnnotationWithoutInput("/scratch/projects/bbc/aiminy_project/7_27_2017_bam/","/scratch/projects/bbc/aiminy_project/macs2_call","hs","macs2",0.0001)'
#
peakCallAndAnnotationWithoutInput <- function(input.file.dir,output.file.dir,genome = "hs",peakcaller = c("macs14", "macs2"), peakPvalue) {
genome <- match.arg(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
PATH1 = Sys.getenv("PATH")
macs2_Lib = file.path("/nethome/axy148/NGS_tools/MACS/bin/")
Sys.setenv(PATH = paste0(macs2_Lib, ":", PATH1))
cmd1 <- Sys.which("macs2")[[1]]
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-m 6,18 --bw=200 -p", peakPvalue, sep = " ")
})
re<-ParserReadFiles(input.file.dir,"bam")
file.name.2<-re$input
#output.dir.name=re$output
temp3=file.path(output.file.dir,"PeakCall")
if(!dir.exists(temp3)){dir.create(temp3,recursive = TRUE)}
re.out<-file.name.2
#cmd9="macs14 -t "
#cmd10="-f BAM -g hs -n "
#cmd11=" -m 6,18 --bw=200 -p 0.00001"
lapply(1:length(re.out),function(u,re.out,temp3){
x=re.out[[u]]
x_name=names(re.out)[u]
cmd12=paste(cmd9,x,cmd10,file.path(temp3,paste0(x_name,"_hs_1.00e-05_macs")),cmd11,sep=" ")
print(cmd12)
system(cmd12, intern = TRUE, ignore.stderr = TRUE)
#re=read.table(u,header=FALSE)
# re<-as.character(re[,1])
# #colnames(re)=c("Count","GeneName")
# re
},re.out,temp3)
#AnnotatePeak2(paste0(temp3,"/"),"*macs142_peaks.bed",7,paste0(output.dir.name,"PeakAnnotation_at_",temp2),genome="Hs")
#AnnotatePeak3(paste0(temp3,"/"),paste0(output.dir.name,"_PeakAnnotation"),
# genome="Hs")
#BamFileSortIndexVisualization2(re,genome)
}
#input <- "~/nodup.bam.txt"
#output <- "~/"
#
# R -e 'library(ChipSeq);library(ChipSeq);ChipSeq:::parseToSampleInfo("/projects/scratch/bbc/Project/Danny_chip3/Filtered_bam","*.bam$","~/Danny_chip3","sample_infor_Danny_chip3.txt","Danny_chip3")'
#
parseToSampleInfo <- function(input.dir,input.pattern,output.dir,output.file,label){
file.1 <- list.files(input.dir,pattern=input.pattern, all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
s <- file.1
ss <- basename(as.character(s))
sss <- tools::file_path_sans_ext(ss)
ssss <- tools::file_path_sans_ext(sss)
sm.info <- cbind.data.frame(sss,rep(label,length(sss)),ssss,rep("Link",length(sss)))
colnames(sm.info) <- c("ID","Type_Cell","Type_TF","Link")
if(!dir.exists(output.dir)){dir.create(output.dir,recursive = TRUE)}
write.table(sm.info,file=file.path(output.dir,output.file),row.names = FALSE,quote=FALSE,sep=",")
}
# ChipSeq:::getAb2inputPair("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt")
#
getAb2inputPair <- function(input.infor.file)
{
f <- read.table(input.infor.file,header = FALSE)
colnames(f)=c("Ab","Input")
print(f)
return(f)
}
# ChipSeq:::getBwUseBamCompare("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt","~")
#
getBwUseBamCompare <- function(input.infor.file,bl_file,output.dir)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
x.run <- apply(x.sample, 1, function(x,output.dir)
{
print(t(x))
x <- as.data.frame(t(x))
xx <- x$Ab
xx.input <- x$Input
#xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd1 <- paste("bamCompare --bamfile1",xx,"--bamfile2",xx.input,sep=" ")
cmd2 <- "--binSize 25"
#cmd3 <- "--ratio log2 -o"
cmd3 <- "--ratio subtract --normalizeTo1x 2451960000 --ignoreForNormalization chrX -o"
cmd3 <- "--ratio subtract --normalizeTo1x 2451960000 --ignoreForNormalization chrX --blackListFileName"
cmd3 <- paste(cmd3,bl_file,"-o",sep=" ")
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("log2ratio_",basename(as.character(xx)),"_vs_",basename(as.character(xx.input)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
print(cmd6)
system(cmd6)
}, output.dir)
}
# R -e 'library(ChipSeq)::peakCall2("/Volumes/Bioinformatics$/Aimin_project/Danny_chip3.txt","hs","~/","macs2",0.00001)'
#
peakCall2 <- function(input.infor.file,genome = c("Hs","hs", "HS", "hS"),output.dir,peakcaller = c("macs14","macs2"), peakPvalue)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
peakcaller <- match.arg(peakcaller)
genome <- match.arg(genome)
genome <- tolower(genome)
cmd10 <- paste("-f BAM", "-g", genome, "-n", sep = " ")
switch(peakcaller, macs2 = {
cmd1 <- "macs2"
cat(cmd1, "\n")
cmd9 = paste(cmd1, "callpeak -t", sep = " ")
cmd11 <- paste("-p", peakPvalue, sep = " ")
}, {
cmd9 = "macs14 -t "
cmd11 <- paste("-p", peakPvalue, sep = " ")
})
macs.run <- apply(x.sample,1,function(u, output.dir)
{
x <- as.data.frame(t(u))
xx <- x$Ab
xx.input <- x$Input
x.name <- paste(peakcaller,basename(as.character(xx)),basename(as.character(xx.input)),genome,peakPvalue,sep="_")
cmd12 = paste(cmd9, xx, "-c", xx.input, cmd10, file.path(output.dir, x.name), cmd11, sep = " ")
print(cmd12)
system(cmd12)
},output.dir)
#AnntationUsingChipSeeker(output.dir, "peaks.bed", temp3, DD = 5000)
# return(re)
}
useBamCoverage <- function(input.sample.file,input.bam.file,output.dir)
{
re <- GetSampleInfo(input.sample.file, input.bam.file)
cellInfo <- re$y
# output.dir.name = dirname(input.sample.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
temp3 = output.dir
# cmd9 = 'ngs.plot.r -G' cmd10 = '-R' cmd11 = '-C' cmd12 = '-O' cmd13 = '-T'
# cmd14 = '-L' cmd15 = '-RR' cmd16 = '-CD' cmd17= '-GO'
cellInfo.run <- lapply(1:length(cellInfo), function(u,cellInfo,
temp3)
{
x.name = cellInfo[[u]]$name
es <- cellInfo[[u]]$es
x.input <- es[es$Type_TF == "Input", ]$file.name
x.sample <- es[es$Type_TF != "Input", ]
#print(x.sample)
#print(x.input)
x.run <- apply(x.sample, 1, function(x, x.input, temp3)
{
y <- x
ID <- y[1]
Type_Cell <- y[2]
Type_TF <- y[3]
Cell_TF <- y[4]
file.name <- y[5]
xx <- file.name
xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
# ~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCompare --bamfile1 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-03_S11_R1.marked_sorted.bam --bamfile2 /scratch/projects/bbc/aiminy_project/DannyNewNgsPlot/2017-03-02-17_S1_R1.marked_sorted.bam --binSize 25 --ratio log2 -o ~/BamCompare/log2ratio_2017-03-02-03_S11_R1.marked_sorted.bam_vs_2017-03-02-17_S1_R1.marked_sorted.bam.bw
#bamCoverage -b reads.bam -o coverage.bw
#bamCoverage --bam a.bam -o a.SeqDepthNorm.bw \
#--binSize 10
#--normalizeTo1x 2150570000
#--ignoreForNormalization chrX
#--extendReads
cmd1 <- paste("~/python/Python-2.7.11/python ~/NGS_tools/deepTools/bin/bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX"
cmd3 <- "-o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("To1x_",basename(as.character(xx)),"_vs_","no_input",".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
#cat(cmd6, "\n")
#cat("\n")
}, x.input, temp3)
x.run
}, cellInfo,temp3)
names(cellInfo.run) = unlist(lapply(cellInfo, function(u)
{
u$name
}))
zzz <- unlist(cellInfo.run)
lapply(1:length(zzz), function(u, zzz)
{
cat(as.character(zzz[u][[1]]), "\n")
cat("\n")
system(as.character(zzz[u][[1]]))
}, zzz)
# # dir.name=temp3 dir.name=reformatPath(dir.name)
#
# file.name = file.path(temp3, dir(temp3, recursive = TRUE))
#
# file.name.2 <- as.list(file.name)
#
#
# # names(file.name.2) = unlist(lapply(file.name.2, function(u) { u$name }))
#
# zzz <- unlist(file.name.2)
#
# lapply(1:length(zzz), function(u, zzz)
# {
#
# dir.name = dirname(zzz[u][[1]])
# file_name = file_path_sans_ext(basename(zzz[u][[1]]))
#
# cmd = paste("ngs.plot.r -G hg19 -R tss -C", zzz[u][[1]], "-O", file.path(dir.name,
# paste0(file_name, ".tss")), "-T", file_name, "-L 4000 -RR 1 -CD 1 -GO total",
# sep = " ")
#
#
#
# # system(as.character(zzz[u][[1]])) job.name = paste0('bamPlot.', u)
# # cmd.pegasus = usePegasus(job.option, Wall.time = '72:00',cores = 32,Memory
# # = 16000,span.ptile = 16,job.name) cmd2 = paste(cmd.pegasus,cmd,sep = ' ')
#
# cmd2 = cmd
# cat(cmd2, "\n")
# cat("\n")
# system(cmd2)
# }, zzz)
#
#
# re <- list(cellInforun = cellInfo.run, zzz = zzz)
#
# # AnntationUsingChipSeeker(temp3, 'peaks.bed', temp3, DD = 5000)
#return(re)
}
getBwUseBamCoverage2 <- function(input.infor.file,bl_file,output.dir)
{
x.sample <- getAb2inputPair(input.infor.file)
if (!dir.exists(output.dir))
{
dir.create(output.dir, recursive = TRUE)
}
x.run <- apply(x.sample, 1, function(x,output.dir)
{
print(t(x))
x <- as.data.frame(t(x))
xx <- x$Ab
xx.input <- x$Input
#xx.name = paste(ID, gsub(" ", "-", Type_Cell), Type_TF, sep = "-")
cmd1 <- paste("bamCoverage --bam",xx,sep=" ")
cmd2 <- "--binSize 25 --normalizeTo1x 2451960000 --ignoreForNormalization chrX --blackListFileName"
cmd2 <- paste(cmd2,bl_file,sep=" ")
cmd3 <- "-o"
#cmd1 <- paste("bamCompare --bamfile1",xx,"--bamfile2",xx.input,sep=" ")
#cmd2 <- "--binSize 25"
#cmd3 <- "--ratio log2 -o"
cmd4 <- paste(cmd1,cmd2,cmd3,sep=" ")
cmd5 <- file.path(output.dir,paste0("Un_norma",basename(as.character(xx)),".bw"))
cmd6 <- paste(cmd4,cmd5,sep=" ")
cmd6
print(cmd6)
system(cmd6)
}, output.dir)
}
getSummitSequence<-function(dir.name,input.file.pattern,genome,out.dir.name){
if (!dir.exists(out.dir.name))
{
dir.create(out.dir.name, recursive = TRUE)
}
file.1 <- list.files(dir.name,pattern=input.file.pattern,all.files = TRUE,full.names = TRUE,recursive = TRUE,include.dirs = TRUE)
print(file.1)
re.out<-lapply(file.1,function(u){
#print(names(u))
uu=u
re=read.table(uu,header=TRUE)
print(head(re))
colnames(re)[c(7,9)]=c("-10*LOG10(pvalue)","FDR(%)")
temp<-re
temp$start<-temp$start-1
temp$end<-temp$end-1
temp$summit<-temp$abs_summit-1
temp
summitPeak<-temp$start+temp$summit
temp2<-temp
temp2$start<-summitPeak-49
temp2$end<-summitPeak+50
temp2$summit<-summitPeak
temp2$length<-temp2$end-temp2$start+1
temp2
rownames(temp2)<-paste0("MACS_peak_",rownames(temp2))
temp3<-toGRanges(temp2)
names(temp3)<-rownames(temp2)
if(genome == "mm10")
{
dd.GRCm39.mm10<-toGRanges(EnsDb.Mmusculus.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Mmusculus.UCSC.mm10")
genome(temp3.trimmed)<-"mm10"
}
if(genome == "hg19"){
dd.GRCm39.mm10<-toGRanges(EnsDb.Hsapiens.v75)
genome(temp3)<-genome(dd.GRCm39.mm10)
seqlevels(dd.GRCm39.mm10,force=TRUE) <- seqlevels(temp3)
seqinfo(temp3)<-seqinfo(dd.GRCm39.mm10)
temp3.trimmed<-trim(temp3, use.names=TRUE)
genome.mm10<-getBSgenome("BSgenome.Hsapiens.UCSC.hg19")
genome(temp3.trimmed)<-"hg19"
}
seq.temp3<-getAllPeakSequence(temp3.trimmed,genome=genome.mm10)
write2FASTA(seq.temp3, file.path(out.dir.name,paste0(basename(u),".fa")))
#re2<-list(originalPeak=re,down1Peak=temp,aroundSummit100Peak=temp2,GR=temp3)
#re2
})
#sample.name<-sapply(strsplit(names(file.name.2),split="_peaks_"),"[[",1)
#names(re.out)<-sample.name
test
#return(re.out)
}
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