# Method: mapInit ####
#' @export
mapInit.DR2S <- function(x, opts = list(), ...) {
x$runMapInit(opts = opts, ...)
invisible(x)
}
DR2S_$set("public", "runMapInit", function(opts = list(), ...) {
# debug
# opts = list()
# library(assertthat)
# library(foreach)
# library(futile.logger)
# library(ggplot2)
# library(cowplot)
# library(S4Vectors)
# library(Rsamtools)
# self <-dr2s
# self <- mapper
flog.info("# mapInit", name = "info")
## Collect starttime for mapInit runstats
start.time <- Sys.time()
## Initiate indenter
indent <- indentation(1)
indent2 <- incr(indent)
## Export mapInit config to function environment
args <- self$getOpts("mapInit")
list2env(args, envir = environment())
assert_that(
exists("includeDeletions") && is.logical(includeDeletions),
exists("includeInsertions") && is.logical(includeInsertions),
exists("callInsertionThreshold") && is.numeric(callInsertionThreshold),
exists("minMapq") && is.numeric(minMapq),
exists("topx"),
exists("pickiness") && is.numeric(pickiness),
exists("increasePickiness") && is.numeric(increasePickiness),
exists("lowerLimit") && is.numeric(lowerLimit),
exists("updateBackgroundModel") && is.logical(updateBackgroundModel),
exists("createIgv") && is.logical(createIgv),
exists("plot") && is.logical(plot)
)
## Get options and prepare mapping
outdir <- .dirCreateIfNotExists(self$absPath("mapInit"))
.dirCreateIfNotExists(file.path(self$absPath(".plots")))
clean <- TRUE
pickedTopX <- FALSE
igv <- list()
SR <- list()
if (self$hasShortreads()) {
SR <- mapInitSR(
self = self, opts = opts, includeDeletions = includeDeletions,
includeInsertions = includeInsertions, callInsertions = TRUE,
callInsertionThreshold = callInsertionThreshold, clip = FALSE,
distributeGaps = FALSE, removeError = TRUE, topx = 0,
outdir = outdir, clean = clean, minMapq = 50, indent = indent, ...)
### TODO wrap this command up
if (createIgv)
igv[["SR"]] <- createIgvJsFiles(
reference = self$absPath(refpath(SR$SR2)),
bamfile = self$absPath(bampath(SR$SR2)),
outdir = outdir,
sampleSize = 100)
reffile <- self$absPath(conspath(SR$SR1))
refname <- consname(SR$SR1)
}
else {
maplabel <- "mapInit1"
mapfun <- self$getLrdMapFun()
readtype <- self$getLrdType()
readfile <- self$getLongreads()
reffile <- self$getRefPath()
refname <- self$getReference()
pileup <- mapReads(
mapfun = mapfun, maplabel = maplabel, reffile = reffile, refname = refname,
readfile = readfile, readtype = readtype, opts = opts, outdir = outdir,
includeDeletions = TRUE, includeInsertions = TRUE, callInsertions = TRUE,
callInsertionThreshold = callInsertionThreshold, clip = FALSE,
distributeGaps = TRUE, removeError = TRUE, topx = topx,
updateBackgroundModel = updateBackgroundModel, clean = clean,
minMapq = minMapq, pickiness = pickiness, lowerLimit = lowerLimit,
indent = indent, ...)
## If topx = "auto" or set to non-zero generate a new subsampled
## read file
if (!is.null(picked1 <- reads(pileup))) {
fqFile <- dot(c(self$getSampleId(), readtype, "n" %<<% length(picked1), "fastq", "gz"))
topxFqPath <- .fileDeleteIfExists(file.path(outdir, strip(fqFile, "_")))
names(topxFqPath) <- self$relPath(topxFqPath)
fqReads <- .extractFastq(bampath(pileup), picked1)
ShortRead::writeFastq(fqReads, topxFqPath, compress = TRUE)
## picking plot 1
plotpath <- file.path(outdir, "plot.readpicking1.png")
cowplot::save_plot(plotpath, attr(picked1, "plot"), dpi = 150)
## set flag
pickedTopX <- TRUE
}
## Construct initial longread consensus sequence
refname <- refname %<<% ".consensus"
self$setRefPath(file.path(outdir, strip(maplabel %<<% "." %<<% refname %<<% ".fa", "_")))
reffile <- self$getRefPath()
flog.info("%sConstruct consensus <%s>", indent2(), names(reffile), name = "info")
conseq <- .writeConseq(x = pileup, name = refname, type = "prob",
threshold = NULL, suppressAllGaps = TRUE,
replaceIndel = "N", conspath = reffile)
}
maplabel <- "mapInit2"
mapfun <- self$getLrdMapFun()
readtype <- self$getLrdType()
indelRate <- if (self$hasShortreads()) indelRate(SR$SR2$pileup) else indelRate(pileup)
if (pickedTopX) {
readfile <- topxFqPath
pickiness <- pickiness/increasePickiness
} else {
readfile <- self$getLongreads()
}
pileup <- mapReads(
mapfun = mapfun, maplabel = maplabel, reffile = reffile, refname = refname,
readfile = readfile, readtype = readtype, opts = opts, outdir = outdir,
includeDeletions = TRUE, includeInsertions = FALSE, callInsertions = FALSE,
clip = FALSE, distributeGaps = TRUE, removeError = TRUE, topx = topx,
updateBackgroundModel = updateBackgroundModel, clean = clean,
minMapq = minMapq, pickiness = pickiness, lowerLimit = lowerLimit,
indent = indent, indelRate = indelRate, ...)
if (!is.null(picked2 <- reads(pileup))) {
## picking plot 2
plotpath <- file.path(outdir, "plot.readpicking2.png")
cowplot::save_plot(plotpath, attr(picked2, "plot"), dpi = 150)
}
if (createIgv)
igv[["LR"]] <- createIgvJsFiles(
reference = refpath(pileup),
bamfile = bampath(pileup),
outdir = outdir,
sampleSize = 100,
fragmentReads = TRUE)
self$mapInit = MapList_(
## mapdata
readpath = self$relPath(readfile),
refpath = self$relPath(reffile),
bampath = self$relPath(bampath(pileup)),
conspath = NULL,
pileup = pileup,
stats = list(coverage = .coverage(pileup)),
## metadata
maplabel = maplabel,
refname = refname,
mapper = self$getLrdMapper(),
opts = opts,
## additional metadata
SR1 = SR$SR1,
SR2 = SR$SR2,
igv = igv)
createIgvConfigs(x = self, map = "mapInit", open = "FALSE")
if (plot) {
flog.info("%sPlot MapInit summary", indent(), name = "info")
## Coverage and frequency of minor alleles
p <- self$plotMapInitSummary(thin = 0.25, width = 2)
plotRows <- ifelse(self$hasShortreads(), 2, 1)
cowplot::save_plot(self$absPath("plot.mapInit.png"),
plot = p, ncol = 1, nrow = plotRows, dpi = 150,
base_aspect_ratio = as.numeric(dot(c(5, plotRows))),
title = dot(c(self$getLocus(), self$getSampleId())))
cowplot::save_plot(self$absPath(".plots/plot.mapInit.svg"),
plot = p, ncol = 1, nrow = plotRows,
base_aspect_ratio = as.numeric(dot(c(5, plotRows))))
}
## set mapInit runstats
if (is(meta(self$mapInit, "SR2"), "MapList")) {
.setRunstats(self, "mapInit",
list(Runtime = format(Sys.time() - start.time),
SRcoverage = stats(meta(self$mapInit, "SR2"), "coverage")[["50%"]],
LRcoverage = stats(self$mapInit, "coverage")[["50%"]]))
} else {
.setRunstats(self, "mapInit",
list(Runtime = format(Sys.time() - start.time),
LRcoverage = stats(self$mapInit, "coverage")[["50%"]]))
}
return(invisible(self))
})
# Method: partitionLongreads ####
#' @export
partitionLongreads.DR2S <- function(x) {
## Collect start time for partitionLongreads runstats
start.time <- Sys.time()
x$runPartitionLongreads()
x$runSplitLongreadsByHaplotype()
x$runExtractPartitionedLongreads()
## set partitionLongreads runstats
.setRunstats(x, "partitionLongreads",
list(Runtime = format(Sys.time() - start.time)))
return(invisible(x))
}
DR2S_$set("public", "runPartitionLongreads", function() {
## debug
# library(futile.logger)
# library(assertthat)
# self <- dr2s
flog.info("# PartitionLongreads", name = "info")
## Initiate indenter
indent <- indentation(1)
## Export partitionLongreads config to function environment
args <- self$getOpts("partitionLongreads")
list2env(args, envir = environment())
assert_that(
self$hasMapInit(),
exists("threshold") && is.double(threshold),
exists("distAlleles") && is.count(distAlleles),
exists("skipGapFreq") && is.numeric(skipGapFreq),
exists("noGapPartitioning") && is.logical(noGapPartitioning),
exists("selectCorrelatedPositions") && is.logical(selectCorrelatedPositions),
exists("measureOfAssociation") && is.character(measureOfAssociation),
exists("proportionOfOverlap") && is.numeric(proportionOfOverlap),
exists("minimumExpectedDifference") && is.numeric(minimumExpectedDifference),
exists("selectAllelesBy") && is.character(selectAllelesBy),
exists("minClusterSize") && is.numeric(minClusterSize),
exists("plot") && is.logical(plot)
)
## Get the reference sequence
if (self$hasShortreads()) {
useSR <- TRUE
flog.info("%sConstruct SNP matrix from shortreads", indent(), name = "info")
} else {
useSR <- FALSE
flog.info("%sConstruct SNP matrix from longreads", indent(), name = "info")
}
ppos <- self$polymorphicPositions(useSR = useSR)
## Spurious gaps, especially in longreads can hinder a correct clustering
## Remove gap positions for clustering
if (noGapPartitioning) {
allpp <- NROW(ppos)
ppos <- dplyr::filter(ppos, a1 != "-" & a2 != "-")
flog.info("%sUse %s non-indel polymorphisms out of %s for clustering", indent(),
NROW(ppos), allpp, name = "info")
}
## Check if already finished because it is a homozygous sample
if (NROW(ppos) == 0) {
flog.warn("%sNo polymorphic positions found for clustering", indent(), name = "info")
flog.info("%sEntering single allele polish and report pipeline", indent(), name = "info")
## set runstats
.setRunstats(self, "partitionLongreads",
list(foundPolymorphicPositions = 0L))
return(invisible(.finishCn1(x = self, plot = plot)))
}
mat <- SNPmatrix(self$absPath(bampath(self$mapInit)), ppos)
base_height <- max(6, floor(sqrt(NCOL(mat))))
spos <- .selectAssociatedPolymorphicPositions(
mat, measureOfAssociation, proportionOfOverlap, minimumExpectedDifference,
noSelect = !selectCorrelatedPositions, indent = indent)
mat0 <- mat[, spos[order(as.numeric(spos))], drop = FALSE]
if (plot) {
## correlogram
plotpath <- file.path(self$getOutdir(), "plot.correlogram.png")
cowplot::save_plot(plotpath, attr(spos, "snp.correlogram"),
base_height = base_height, dpi = 150)
## associiation plot
plotpath <- file.path(self$getOutdir(), "plot.association.png")
cowplot::save_plot(plotpath, attr(spos, "snp.association"),
base_height = base_height/2.4, dpi = 150,
base_aspect_ratio = 3)
## if exists: cluster overlap
if (has_attr(spos, "ovl.plot")) {
plotpath <- file.path(self$getOutdir(), "plot.mclust.ovl.png")
grDevices::png(filename = plotpath, width = 5, height = 4.25, units = "in",
res = 150, bg = "white")
print(attr(spos, "ovl.plot"))
grDevices::dev.off()
}
## if exists: mclust
if (has_attr(spos, "mclust")) {
plotpath <- file.path(self$getOutdir(), "plot.mclust.png")
p <- factoextra::fviz_mclust(attr(spos, "mclust"))
cowplot::save_plot(plotpath, p, base_height = 5, dpi = 150)
}
}
flog.info("%sPartition %s longreads over %s SNPs", indent(), NROW(mat0), NCOL(mat0), name = "info")
prt <- partitionReads(x = mat0,
skipGapFreq = skipGapFreq,
deepSplit = 1,
threshold = threshold,
distAlleles = distAlleles,
selectAllelesBy = selectAllelesBy,
minClusterSize = minClusterSize,
indent = incr(indent))
if (plot) {
ppos <- SNP(prt)
## ggseqlogo doesn't work with 1-column matrices
if (length(ppos) > 1) {
pwm <- lapply(PWM(prt), function(pwm) {
pwm[pwm < 0.1] <- 0
pwm
})
p <- suppressMessages(self$plotSeqLogo(ppos, pwm))
cowplot::save_plot(filename = self$absPath("plot.sequence.png"),
plot = p, dpi = 150, units = "cm", limitsize = FALSE,
base_width = 0.4*length(ppos) + 1.4,
base_height = 2.5*length(pwm),
title = dot(c(self$getLocus(), self$getSampleId())))
cowplot::save_plot(filename = self$absPath(".plots/plot.sequence.svg"),
plot = p, units = "cm", limitsize = FALSE,
base_width = 0.4*length(ppos) + 1.4,
base_height = 2.5*length(pwm))
}
}
## set runstats
.setRunstats(self, "partitionLongreads",
list(nLongreads = NROW(mat),
foundPolymorphicPositions = NCOL(mat),
usedPolymorphicPositions = K(prt),
foundClades = OC(prt),
usedClades = as.list(table(PRT(prt)))))
## Set sample haplotypes
self$setHapTypes(levels(as.factor(PRT(prt))))
# Check if we have only one cluster and finish the pipeline if so
if (length(self$getHapTypes()) == 1) {
flog.warn("%sOnly one allele left", indent(), name = "info")
flog.info("%sEntering single allele polish and report pipeline", indent(), name = "info")
return(invisible(.finishCn1(x = self, plot = plot)))
}
self$lrpartition = structure(list(
mat = mat,
prt = prt,
hpl = NULL,
lmt = NULL
),
class = c("PartList", "list"))
return(invisible(self))
})
#' @export
print.PartList <- function(x, ...) {
msg <- sprintf("An object of class '%s'.\n", class(x)[1])
msg <- sprintf("%s [Matrix] %s reads; %s polymorphic positions\n",
msg, NROW(x$mat), NCOL(x$mat))
cat(msg)
cat(" [Partition] ")
print(x$prt, ncols = 4, nrows = 6)
cat(" [Haplotypes] ")
print(x$hpl)
}
DR2S_$set("public", "runSplitLongreadsByHaplotype", function() {
## If reporting is already done exit safely
if (.checkReportStatus(self)) return(invisible(self))
assert_that(self$hasLrdPartition())
## Initiate indenter
indent <- indentation(1)
flog.info("%sSplit partitioned longreads by score", indent(), name = "info")
## Export partitionLongreads config to function environment
args <- self$getOpts("partitionLongreads")
list2env(args, envir = environment())
assert_that(
exists("pickiness") && is.numeric(pickiness),
exists("lowerLimit") && is.numeric(lowerLimit),
exists("plot") && is.logical(plot)
)
prt <- partition(self$getPartition())
haplotypes <- levels(prt$haplotype)
tag <- self$getMapTag("init", "LR")
# Set all limits to NULL
self$setLimits(rep(NA, length(haplotypes)))
prts <- lapply(haplotypes, function(hp) dplyr::filter(prt, .data$haplotype == hp))
names(prts) <- haplotypes
scores <- lapply(prts, function(x) x$mcoef)
lmts <- .optimalPartitionLimits(scores, pickiness, lowerLimit)
self$setLimits(setNames(as.list(lmts$limits$score), haplotypes))
self$lrpartition$lmt <- lmts$plt
# Get only reads within the limit
reads <- stats::setNames(lapply(names(self$getLimits()), function(x) {
dplyr::filter(prt, .data$haplotype == x, .data$mcoef >= self$getLimits()[x])
}), names(self$getLimits()))
## Initiate indenter
indent2 <- incr(indent)
for (hp in haplotypes) {
flog.info("%s<%s>: Using %s longreads with score > %.2f",
indent2(), hp, nrow(reads[[hp]]), self$getLimits()[hp],
name = "info")
}
## set runstats
.setRunstats(self, "partitionLongreads",
list(reads = setNames(as.list(as.integer(lmts$limits$nreads)), haplotypes)))
# results structure
self$lrpartition$hpl <- structure(
setNames(lapply(reads, function(x) {
structure(
x$read,
q = x$mcoef,
freq = NROW(x)/NROW(dplyr::bind_rows(reads)),
limit = self$getLimits()[[x$haplotype[1]]]
)}), haplotypes),
class = c("HapList", "list"))
if (plot) {
## Plot the consensus sequences from clustering
.browseSeqs(SQS(self$getPartition()),
file = self$absPath("partition.fa.html"),
openURL = FALSE)
p <- self$plotPartitionSummary(
label = tag, limits = unlist(self$getLimits()))
cowplot::save_plot(self$absPath("plot.partition.png"), plot = p, dpi = 150,
base_height = 12, base_width = 10,
title = dot(c(self$getLocus(), self$getSampleId())))
cowplot::save_plot(self$absPath(".plots/plot.partition.svg"), plot = p,
base_aspect_ratio = 1.2)
}
return(invisible(self))
})
#' @export
print.HapList <- function(x, ...) {
msg <- sprintf("An object of class '%s'.\n", class(x)[1])
for (haplotype in names(x)) {
msg <- msg %<<%
sprintf("%s: n %s; frequency %s; limit %s\n",
haplotype,
length(x[haplotype]),
round(attr(x[[haplotype]], "freq"), 3),
attr(x[[haplotype]], "limit"))
}
cat(msg)
}
DR2S_$set("public", "runExtractPartitionedLongreads", function() {
## Check if reporting has already been done and exit safely
if (.checkReportStatus(self)) return(invisible(self))
assert_that(self$hasHapList())
## Initiate indenter
indent <- indentation(1)
## Export PartitionLongreadsconfig to function environment
args <- self$getOpts("partitionLongreads")
list2env(args, envir = environment())
## extract msa from mapInit
bamfile <- self$absPath(bampath(self$mapInit))
mat <- .msaFromBam(Rsamtools::BamFile(bamfile), paddingLetter = ".")
flog.info("%sExtract partitioned longreads and construct consensus" %<<%
" sequences based on <%s>", indent(), names(bamfile), name = "info")
## For each <hptype>:
## create the "mapIter/<hptype>" directory to hold the reads assigned to <hptype>
## extract the reads assigned to <hptype> from the LR bam file from <mapInit>
## construct a consensus sequence from the reads for each <hptype>
## create a <mapIter0> object for each <hptype>
## hptype <- "A"
## hptype <- "B
indent2 <- incr(indent)
for (hptype in self$getHapTypes()) {
flog.info("%sFor haplotype <%s>:", indent(), hptype, name = "info")
## create the "mapIter/<hptype>" directory
fqdir <- .dirCreateIfNotExists(
normalizePath(file.path(self$getOutdir(), "mapIter", hptype), mustWork = FALSE))
## get the ids of the reads assigned to <hptype>
readIds <- self$getHapList(hptype)
## extract these reads and write to file
fq <- .extractFastq(bamfile, qnames = readIds)
fqfile <- dot(c(
"hap" %<<% hptype, self$getLrdType(), self$getLrdMapper(),
"n" %<<% length(fq), "fastq", "gz"))
fqout <- .fileDeleteIfExists(file.path(fqdir, fqfile))
nfq <- ShortRead::writeFastq(fq, file = fqout, full = FALSE, compress = TRUE)
## assert that all records got written to file
assert_that(nfq == length(fq))
## Extract consensus matrix from mapInit for the clustered reads
cmat <- .extractIdsFromMat(mat, readIds)
## Construct the initial longread consensus sequence for <hptype>
maplabel <- "mapIter0"
consname <- maplabel %<<% ".consensus." %<<% hptype
conspath <- file.path(fqdir, consname %<<% ".fa")
names(conspath) <- self$relPath(conspath)
flog.info("%sConstruct consensus <%s>", indent2(), names(conspath), name = "info")
conseq <- .writeConseq(x = cmat, name = consname, type = "prob",
threshold = NULL, suppressAllGaps = TRUE,
replaceIndel = "", conspath = conspath)
##
self$mapIter$`0`[[hptype]] = MapList_(
## mapdata
readpath = self$relPath(fqout),
refpath = refpath(self$mapInit),
bampath = bampath(self$mapInit),
conspath = self$relPath(conspath),
pileup = self$mapInit$pileup,
stats = list(coverage = setNames(nfq, "50%")),
## required metadata
maplabel = maplabel,
refname = refname(self$mapInit),
mapper = meta(self$mapInit, "mapper"),
opts = meta(self$mapInit, "opts")
)
}
return(invisible(self))
})
## Method: mapIter ####
#' @export
mapIter.DR2S <- function(x, opts = list(), ...) {
x$runMapIter(opts = opts, ...)
return(invisible(x))
}
DR2S_$set("public", "runMapIter", function(opts = list(), ...) {
# debug
# self <- dr2s
# opts = list()
## If reporting is already done exit safely
if (.checkReportStatus(self)) return(invisible(self))
## Collect start time for mapIter runstats
start.time <- Sys.time()
flog.info("# MapIter", name = "info")
## Initiate indenter
indent <- indentation(1)
flog.info("%sIterative mapping of partitioned longreads", indent(), name = "info")
## Export mapIter config to function environment
args <- self$getOpts("mapIter")
list2env(args, envir = environment())
assert_that(
exists("iterations") && is.count(iterations),
exists("columnOccupancy") && is.double(columnOccupancy),
exists("callInsertionThreshold") && is.double(callInsertionThreshold),
exists("plot") && is.logical(plot)
)
baseoutdir <- self$absPath("mapIter")
clean <- TRUE
includeInsertions <- ifelse(self$hasShortreads(), FALSE, TRUE)
callInsertions <- ifelse(self$hasShortreads(), FALSE, TRUE)
## Mapper
mapfun <- self$getLrdMapFun()
# iteration <- 1
# iteration <- 2
for (iteration in seq_len(iterations)) {
flog.info("%sIteration %s of %s", indent(), iteration, iterations, name = "info")
iterationC <- toString(iteration)
maplabel <- "mapIter" %<<% iterationC
prevIteration <- self$mapIter[[toString(iteration - 1)]]
# hptype = "A"
# hptype = "B"
indent2 <- incr(indent)
foreach(hptype = self$getHapTypes()) %do% {
flog.info("%sFor haplotype <%s>:", indent2(), hptype, name = "info")
readtype <- self$getLrdType()
readfile <- self$absPath(readpath(prevIteration[[hptype]]))
reffile <- self$absPath(conspath(prevIteration[[hptype]]))
refname <- consname(prevIteration[[hptype]])
outdir <- file.path(baseoutdir, hptype)
pileup <- mapReads(
mapfun = mapfun, maplabel = maplabel, reffile = reffile, refname = refname,
readfile = readfile, readtype = readtype, opts = opts, outdir = outdir,
includeDeletions = TRUE, includeInsertions = includeInsertions,
callInsertions = callInsertions, callInsertionThreshold = callInsertionThreshold,
clip = FALSE, distributeGaps = TRUE, removeError = TRUE, topx = 0,
clean = clean, indent = incr(indent2), ...)
## Construct consensus sequence
consname <- maplabel %<<% ".consensus." %<<% hptype
conspath <- file.path(outdir, consname %<<% ".fa")
names(conspath) <- self$relPath(conspath)
flog.info("%sConstruct consensus <%s>", indent2(), names(conspath), name = "info")
conseq <- .writeConseq(x = pileup, name = consname, type = "prob",
threshold = NULL, suppressAllGaps = FALSE,
suppressInsGaps = TRUE, columnOccupancy = columnOccupancy,
replaceIndel = "", conspath = conspath)
## Initialize mapIter MapList
self$mapIter[[iterationC]][[hptype]] = MapList_(
## mapdata
readpath = self$relPath(readfile),
refpath = self$relPath(reffile),
bampath = self$relPath(bampath(pileup)),
conspath = self$relPath(conspath),
pileup = pileup,
stats = list(coverage = .coverage(pileup)),
## required metadata
maplabel = maplabel,
refname = refname,
mapper = self$getLrdMapper(),
opts = opts
)
}
}
if (plot) {
flog.info("%sPlot MapIter summary", indent(), name = "info")
## Coverage and base frequency
plotlist <- foreach(iteration = seq_len(self$getIterations())) %do% {
suppressWarnings(self$plotMapIterSummary(thin = 0.1, width = 4,
iteration = iteration, drop.indels = TRUE))
}
p <- cowplot::plot_grid(plotlist = plotlist, nrow = self$getIterations())
cowplot::save_plot(p, dpi = 150, filename = self$absPath("plot.mapIter.png"),
base_width = 12*length(self$getHapTypes()),
base_height = 3*self$getIterations(),
title = dot(c(self$getLocus(), self$getSampleId())))
cowplot::save_plot(p, filename = self$absPath(".plots/plot.mapIter.svg"),
base_width = 12*length(self$getHapTypes()),
base_height = 3*self$getIterations())
}
createIgvConfigs(x = self, map = "mapIter", open = "FALSE")
## set mapIter runstats
.setRunstats(self, "mapIter",
list(Runtime = format(Sys.time() - start.time)))
return(invisible(self))
})
## Method: partitionShortreads ####
#' @export
partitionShortreads.DR2S <- function(x, opts = list(), ...) {
x$runPartitionShortreads(opts = opts, ...)
invisible(x)
}
# TODO: look at arguments and make same
DR2S_$set("public", "runPartitionShortreads", function(opts = list(), ...) {
## debug
# opts = list()
## If reporting is already done exit safely
if (.checkReportStatus(self)) return(invisible(self))
## Collect start time for partitionShortreads runstats
start.time <- Sys.time()
flog.info("# PartitionShortreads ...", name = "info")
## Initiate indenter
indent <- indentation(1)
## exit savely if shortreads not provided
if (!self$hasShortreads()) {
flog.warn("%sCannot partition shortreads. No shortreads provided", indent(), name = "info")
return(invisible(self))
}
## exit savely if initial SR mapping not performed
if (is.null(meta(self$mapInit, "SR2"))) {
flog.warn("%sCannot partition shortreads. Run 'mapInit()' first", indent(), name = "info")
return(invisible(self))
}
flog.info("%sPartition shortreads based on initial mapping and " %<<%
"longread clustering", indent(), name = "info")
## Export partitionShortreads config to function environment
args <- self$getOpts("partitionShortreads")
list2env(args, envir = environment())
bamfile <- self$absPath(bampath(meta(self$mapInit, "SR2")))
hptypes <- self$getHapTypes()
prtMat <- self$lrpartition$mat
seqs <- lapply(self$lrpartition$hpl, function(x) {
.getSeqsFromMat(as.matrix(prtMat[x, ]))
})
mats <- lapply(seqs, function(x, names) {
magrittr::set_colnames(createPWM(x), names)
}, names = colnames(prtMat))
# Run partitioning
srpartition <- getSRPartitionScores(bamfile, mats)
## Assign read to haplotype with highest probability,
## i.e. product over probabilities of each haplotype and choose max
flog.info("%sGet highest-scoring haplotype for each read", indent(), name = "info")
srpartition$haplotypes <- scoreHighestSR(srpartition$srpartition, diffThreshold = 0.001)
# Write fastqs
# hp <- "A"
indent2 <- incr(indent)
foreach(hp = hptypes) %do% {
srfilenames <- c()
flog.info("%sWrite shortread fastq for haplotype <%s>", indent2(), hp, name = "info")
fqs <- self$getShortreads()
dontUse <- dplyr::filter(srpartition$haplotypes, haplotype != hp)$read
# fq <- fqs[1]
foreach(fq = fqs) %do% {
fqPart <- self$absPath(
file.path(
dirname(readpath(self$mapIter$`0`[[hp]])),
dot(c(strsplit1(basename(fq), ".", fixed = TRUE)[1], hp, "fastq.gz"))
))
.writePartFq(fq, fqPart, dontUse = dontUse, indent = incr(indent2))
srfilenames <- c(srfilenames, fqPart)
self$srpartition[[hp]]$srpartition <- srpartition
NULL
}
self$srpartition[[hp]]$SR <- self$relPath(srfilenames)
NULL
}
## set partitionShortreads runstats
## set mapIter runstats
.setRunstats(self, "partitionShortreads",
list(Runtime = format(Sys.time() - start.time)))
return(invisible(self))
})
## Method: mapFinal ####
#' @export
mapFinal.DR2S <- function(x, opts = list(), ...) {
x$runMapFinal(opts = opts, ...)
invisible(x)
}
DR2S_$set("public", "runMapFinal", function(opts = list(), ...) {
## debug
# opts = list()
# library(futile.logger)
# library(foreach)
# self <- dr2s
## If reporting is already done exit safely
if (.checkReportStatus(self)) return(invisible(self))
## Collect start time for mapFinal runstats
start.time <- Sys.time()
flog.info("# mapFinal", name = "info")
## Initiate indenter
indent <- indentation(1)
flog.info("%sMap longreads and shortreads against mapIter consensus sequences", indent(), name = "info")
## Export mapFinal config to function environment
args <- self$getOpts("mapFinal")
list2env(args, envir = environment())
assert_that(
exists("includeDeletions") && is.logical(includeDeletions),
exists("includeInsertions") && is.logical(includeInsertions),
exists("callInsertionThreshold") && is.numeric(callInsertionThreshold),
exists("trimPolymorphicEnds") && is.logical(trimPolymorphicEnds),
exists("createIgv") && is.logical(createIgv),
exists("plot") && is.logical(plot)
)
igv <- list()
maplabel <- "mapFinal"
outdir <- .dirCreateIfNotExists(self$absPath(maplabel))
lastIter <- self$mapIter[[max(names(self$mapIter))]]
hptypes <- self$getHapTypes()
reffiles <- stats::setNames(lapply(hptypes, function(x)
self$absPath(conspath(lastIter[[x]]))), hptypes)
readfilesLR <- stats::setNames(lapply(hptypes, function(x)
self$absPath(readpath(lastIter[[x]]))), hptypes)
if (self$hasShortreads()) {
readfilesSR <- stats::setNames(lapply(hptypes, function(x)
self$absPath(self$srpartition[[x]]$SR)), hptypes)
}
## hp = "A"
## hp = "B"
for (hp in hptypes) {
flog.info("%sFor haplotype <%s>", indent(), hp, name = "info" )
reffile <- reffiles[[hp]]
##
## [1] Map longreads
##
refname <- "LR" %<<% hp
mapfun <- self$getLrdMapFun()
readfile <- readfilesLR[[hp]]
readtype <- self$getLrdType()
pileup <- mapReads(
mapfun = mapfun, maplabel = maplabel, reffile = reffile, refname = refname,
readfile = readfile, readtype = readtype, opts = opts, outdir = outdir,
includeDeletions = includeDeletions, includeInsertions = includeInsertions,
callInsertions = FALSE, callInsertionThreshold = callInsertionThreshold,
clip = FALSE, distributeGaps = TRUE, removeError = TRUE, topx = 0,
clean = TRUE, max_depth = 1e4, min_mapq = 0, indent = incr(indent), ...)
## Create igv
if (createIgv) {
igv <- createIgvJsFiles(
refpath(pileup), bampath(pileup), self$getOutdir(), sampleSize = 100,
fragmentReads = TRUE)
}
## Construct consensus sequence
consname <- maplabel %<<% ".consensus." %<<% refname
conspath <- file.path(outdir, consname %<<% ".fa")
names(conspath) <- self$relPath(conspath)
flog.info("%sConstruct consensus <%s>", indent(), names(conspath), name = "info")
conseq <- .writeConseq(x = pileup, name = consname, type = "ambig",
threshold = 1/4, suppressAllGaps = TRUE,
replaceIndel = "", conspath = conspath)
## Initialize mapFinal LR MapList
self$mapFinal$LR[[hp]] = MapList_(
## mapdata
readpath = self$relPath(readfile),
refpath = self$relPath(reffile),
bampath = self$relPath(bampath(pileup)),
conspath = self$relPath(conspath),
pileup = pileup,
stats = list(coverage = .coverage(pileup)),
## required metadata
maplabel = maplabel,
refname = refname,
mapper = self$getLrdMapper(),
opts = opts,
## additional metadata
igv = igv
)
if (self$hasShortreads()) {
##
## [2] Map shortreads
##
refname <- "SR" %<<% hp
mapfun <- self$getSrdMapFun()
readfile <- readfilesSR[[hp]]
readtype <- self$getSrdType()
pileup <- mapReads(
mapfun = mapfun, maplabel = maplabel, reffile = reffile, refname = refname,
readfile = readfile, readtype = readtype, opts = opts, outdir = outdir,
includeDeletions = includeDeletions, includeInsertions = includeInsertions,
callInsertions = TRUE, callInsertionThreshold = callInsertionThreshold,
clip = trimPolymorphicEnds, distributeGaps = TRUE, removeError = TRUE, topx = 0,
clean = TRUE, max_depth = 1e5, min_mapq = 50, min_base_quality = 13,
indent = incr(indent), ...)
## Create igv
if (createIgv) {
igv <- createIgvJsFiles(
refpath(pileup), bampath(pileup), self$getOutdir(), sampleSize = 100)
}
## Construct consensus sequence
consname <- maplabel %<<% ".consensus." %<<% refname
conspath <- file.path(outdir, consname %<<% ".fa")
names(conspath) <- self$relPath(conspath)
flog.info("%sConstruct consensus <%s>", indent(), names(conspath), name = "info")
conseq <- .writeConseq(x = pileup, name = consname, type = "ambig",
threshold = 1/4, suppressAllGaps = TRUE,
replaceIndel = "", conspath = conspath)
## Initialize mapFinal SR MapList
self$mapFinal$SR[[hp]] = MapList_(
## mapdata
readpath = self$relPath(readfile),
refpath = self$relPath(reffile),
bampath = self$relPath(bampath(pileup)),
conspath = self$relPath(conspath),
pileup = pileup,
stats = list(coverage = .coverage(pileup)),
## required metadata
maplabel = maplabel,
refname = refname,
mapper = self$getSrdMapper(),
opts = opts,
## additional metadata
igv = igv
)
}
}
if (plot) {
flog.info("%sPlot MapFinal summary", indent(), name = "info")
## Coverage and base frequency
readtypes <- if (self$hasShortreads()) c("LR", "SR") else "LR"
plotRows <- if (self$hasShortreads()) 2 else 1
## readtype = "LR"
plotlist <- foreach(readtype = readtypes) %do% {
suppressWarnings(self$plotMapFinalSummary(readtype = readtype, thin = 0.25, width = 20))
}
p <- cowplot::plot_grid(plotlist = plotlist, nrow = plotRows, labels = readtypes, hjust = -0.25)
cowplot::save_plot(p, dpi = 150, filename = self$absPath("plot.mapFinal.png"),
base_width = 6*plotRows*length(hptypes),
base_height = 6/plotRows*length(readtypes),
title = dot(c(self$getLocus(), self$getSampleId())))
cowplot::save_plot(p, filename = self$absPath(".plots/plot.mapFinal.svg"),
base_width = 6*plotRows*length(hptypes),
base_height = 6/plotRows*length(readtypes))
}
## set mapFinal runstats
.setRunstats(self, "mapFinal",
list(Runtime = format(Sys.time() - start.time)))
return(invisible(self))
})
## Method: runPipeline ####
DR2S_$set("public", "runPipeline", function() {
steps_ <- self$getPipeline()
## Collect start time for DR2Spipeline runstats
start.time <- Sys.time()
while (length(steps_) > 0) {
step <- steps_[1]
self$run_(step)
steps_ <- steps_[-1]
}
## set DR2Spipeline runstats
.setRunstats(self, "DR2Spipeline",
list(Runtime = format(Sys.time() - start.time)))
return(invisible(self))
})
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