#' @title Running RSEM, Li and Dewey BMC Bioinformatics 2011 12:323
#' @description This function executes the docker container rsemstar1, where RSEM is used to calculate gene/isoforms counts using as mapper STAR, Dubin et al. Bioinformatics. 2013 Jan 1;29(1):15-21
#'
#' @param group, a character string. Two options: \code{"sudo"} or \code{"docker"}, depending to which group the user belongs
#' @param fastq.folder, a character string indicating where gzip fastq files are located
#' @param scratch.folder, a character string indicating the scratch folder where docker container will be mounted
#' @param genome.folder, a character string indicating the folder where the indexed reference genome for STAR is located. IMPORTANT the present function only suport genomic indexes made using ensembl genom and the corresponding gtf
#' @param seq.type, a character string indicating the type of reads to be trimmed. Two options: \code{"se"} or \code{"pe"} respectively for single end and pair end sequencing
#' @param strandness, a character string indicating the type ofsequencing protocol used for the analysis. Three options: \code{"none"}, \code{"forward"}, \code{"reverse"} respectively for non strand selection, forward for Illumina strandness protocols, reverse for ACCESS Illumina protocol
#' @param threads, a number indicating the number of cores to be used from the application
#' @param save.bam, a boolean TRUE FALSE to decide if bam files are saved
#' @author Raffaele Calogero
#'
#' @return three files: dedup_reads.bam, which is sorted and duplicates marked bam file, dedup_reads.bai, which is the index of the dedup_reads.bam, and dedup_reads.stats, which provides mapping statistics
#' @examples
#'\dontrun{
#' #downloading fastq files
#' system("wget http://130.192.119.59/public/test_R1.fastq.gz")
#' system("wget http://130.192.119.59/public/test_R2.fastq.gz")
#' library(docker4seq)
#' #running rsemstar nostrand pe
#' rsemstar(group="docker",fastq.folder=getwd(), scratch.folder="/data/scratch/",
#' genome.folder="/data/genomes/hg38star/", seq.type="pe", strandness="none",
#' threads=8, save.bam = FALSE)
#'
#' }
#' @export
rsemstar <- function(group=c("sudo","docker"),fastq.folder=getwd(), scratch.folder="/data/scratch", genome.folder, seq.type=c("se","pe"), strandness=c("none","forward","reverse"), threads=1, save.bam = TRUE){
home <- getwd()
setwd(fastq.folder)
#initialize status
system("echo 0 > ExitStatusFile 2>&1")
#running time 1
ptm <- proc.time()
#running time 1
test <- dockerTest()
if(!test){
cat("\nERROR: Docker seems not to be installed in your system\n")
system("echo 10 > ExitStatusFile 2>&1")
setwd(home)
return(10)
}
#########check scratch folder exist###########
if (!file.exists(scratch.folder)){
cat(paste("\nIt seems that the ",scratch.folder, "folder does not exist\n"))
system("echo 3 > ExitStatusFile 2>&1")
setwd(home)
return(3)
}
#############################################
tmp.folder <- gsub(":","-",gsub(" ","-",date()))
scrat_tmp.folder=file.path(scratch.folder, tmp.folder)
writeLines(scrat_tmp.folder,paste(fastq.folder,"/tempFolderID", sep=""))
cat("\ncreating a folder in scratch folder\n")
dir.create(file.path(scratch.folder, tmp.folder))
dir.create(file.path(scratch.folder, tmp.folder,"/tmp"))
dir <- dir(path=fastq.folder)
dir.info <- dir[which(dir=="run.info")]
if(length(dir.info)>0){
system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
system(paste("cp ",fastq.folder,"/run.info ", scratch.folder,"/",tmp.folder,"/run.info", sep=""))
}
dir <- dir[grep(".fastq.gz$", dir)]
dir.trim <- dir[grep("trimmed", dir)]
cat("\ncopying \n")
if(length(dir)==0){
cat(paste("It seems that in ", fastq.folder, "there are not fastq.gz files"))
system("echo 1 > ExitStatusFile 2>&1")
return(1)
}else if(length(dir.trim)>0){
dir <- dir.trim
system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
for(i in dir){
system(paste("cp ",fastq.folder,"/",i, " ",scratch.folder,"/",tmp.folder,"/",i, sep=""))
}
system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
}else if(length(dir)>2){
cat(paste("It seems that in ", fastq.folder, "there are more than two fastq.gz files"))
system("echo 2 > ExitStatusFile 2>&1")
setwd(home)
return(2)
}else{
system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
for(i in dir){
system(paste("cp ",fastq.folder,"/",i, " ",scratch.folder,"/",tmp.folder,"/",i, sep=""))
}
system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
}
#Trimmed fastq linking fpr docker
docker_fastq.folder=file.path("/data/scratch", tmp.folder)
#Trimmed fastq linking fpr docker
fastq <- sub(".gz$", "", dir)
cat("\nsetting as working dir the scratch folder and running docker container\n")
if(group=="sudo"){
# system("sudo docker pull docker.io/repbioinfo/rsemstar.2017.01")
if(seq.type=="pe"){
if(strandness=="none"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_nostrand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}else if(strandness=="forward"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_strand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}else if(strandness=="reverse"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_access.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}
}else{
if(strandness=="none"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_nostrand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}else if(strandness=="forward"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_strand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}else if(strandness=="reverse"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_access.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="sudo", params=params)
}
}
}else{
if(seq.type=="pe"){
if(strandness=="none"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_nostrand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}else if(strandness=="forward"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_strand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}else if(strandness=="reverse"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem2_access.sh ",docker_fastq.folder," ", threads," ", fastq[1]," ", fastq[2]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}
}else{
if(strandness=="none"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_nostrand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}else if(strandness=="forward"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_strand.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}else if(strandness=="reverse"){
params <- paste("--cidfile ",fastq.folder,"/dockerID -v ",scratch.folder,":/data/scratch -v ",genome.folder,":/data/genome -d docker.io/repbioinfo/rsemstar.2017.01 sh /bin/rsem1_access.sh ",docker_fastq.folder," ", threads," ", fastq[1]," /data/genome ", fastq.folder, sep="")
resultRun <- runDocker(group="docker", params=params)
}
}
}
if(resultRun==0){
cat("\nThe RSEM STAR analysis is finished\n")
}
#star logs
star.tmp <- dir(paste(file.path(scratch.folder, tmp.folder),"/xxx.temp/", sep=""))
star.tmp <- star.tmp[grep("Log.final.out",star.tmp)]
if(length(star.tmp)>0){
star <- "Log.final.out"
system(paste("cp ",file.path(scratch.folder, tmp.folder),"/xxx.temp/xxxLog.final.out ", file.path(scratch.folder, tmp.folder),"/Log.final.out",sep=""))
}
#system(paste("chmod 777 -R", file.path(scratch.folder, tmp.folder)))
con <- file(paste(file.path(scratch.folder, tmp.folder),"out.info", sep="/"), "r")
tmp <- readLines(con)
close(con)
for(i in tmp){
i <- sub("mv ",paste("mv ",file.path(scratch.folder, tmp.folder),"/",sep=""),i)
system(i)
}
#running time 2
ptm <- proc.time() - ptm
con <- file(paste(fastq.folder,"run.info", sep="/"), "r")
tmp.run <- readLines(con)
close(con)
tmp.run[length(tmp.run)+1] <- paste("RSEM user run time mins ",ptm[1]/60, sep="")
tmp.run[length(tmp.run)+1] <- paste("RSEM system run time mins ",ptm[2]/60, sep="")
tmp.run[length(tmp.run)+1] <- paste("RSEM elapsed run time mins ",ptm[3]/60, sep="")
writeLines(tmp.run,paste(fastq.folder,"run.info", sep="/"))
#running time 2
if (save.bam) {
system(paste("cp ", file.path(scratch.folder, tmp.folder),
"/xxx.temp/xxx.bam ", file.path(fastq.folder), "/Aligned.out.bam",
sep = ""))
system(paste("cp ", file.path(scratch.folder, tmp.folder),
"/xxx.transcript.bam ", file.path(fastq.folder),
"/Aligned.toTranscriptome.out.bam", sep = ""))
}
#saving log and removing docker container
container.id <- readLines(paste(fastq.folder,"/dockerID", sep=""), warn = FALSE)
system(paste("docker logs ", container.id, " >& ", "rsemstar_",substr(container.id,1,12),".log", sep=""))
system(paste("docker rm ", container.id, sep=""))
#removing temporary folder
cat("\n\nRemoving the rsemStar temporary file ....\n")
system(paste("rm -R ",scrat_tmp.folder))
system(paste("rm ",fastq.folder,"/dockerID", sep=""))
system(paste("rm ",fastq.folder,"/tempFolderID", sep=""))
#removing temporary folder
system(paste("cp ",paste(path.package(package="docker4seq"),"containers/containers.txt",sep="/")," ",fastq.folder, sep=""))
setwd(home)
}
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