Updated code to work with derfinder 1.0.8 (current release), build pr…

…oper models for run2-v0.0.42 (initial file had models for a different analysis), link to BioC website instead of GitHub repo, and added a short description of files in the README

README.md

@@ -1,5 +1,11 @@
 # LIBD n36 project
 
-This repository is for hosting the code for the LIDB n36 project being undertaken by Andrew Jaffe's lab.
+This repository is for hosting the code for the LIDB n36 project lead by AE Jaffe.
 
 If you have any questions about the files in this directory contact AE Jaffe <[email protected]>.
+
+Scripts use [derfinder](http://www.bioconductor.org/packages/release/bioc/html/derfinder.html) and are organized in the following way:
+
+* [derCoverageInfo](derCoverageInfo/): contains scripts for processing data from BAM files, filtering and creating Rdata files.
+* [fullCoverage](fullCoverage/): similar as _derCoverageInfo_ but without filtering data.
+* [derAnalysis](derAnalysis/): has scripts for creating the models, running the derfinder analysis one chromosome at a time, merging the results, and creating a html report.

derAnalysis/der2Analysis.sh

@@ -1,7 +1,7 @@
 #!/bin/sh
 
 ## Usage
-# sh der2Analysis.sh run1-v0.0.46
+# sh der2Analysis.sh run2-v0.0.42
 
 # Directories
 MAINDIR=/dcs01/lieber/ajaffe/Brain/derRuns/libd_n36

derAnalysis/der2Merge.sh

@@ -1,7 +1,9 @@
 #!/bin/sh
 
 ## Usage
-# sh der2Merge.sh run1-v0.0.46
+# sh der2Merge.sh run2-v0.0.42
+
+## derfinder available at http://www.bioconductor.org/packages/release/bioc/html/derfinder.html
 
 # Directories
 MAINDIR=/dcs01/lieber/ajaffe/Brain/derRuns/libd_n36
@@ -16,21 +18,26 @@ outdir="${PREFIX}"
 sname="${SHORT}.${PREFIX}"
 echo "Creating script ${sname}"
 cat > ${WDIR}/.${sname}.sh <<EOF
-#!/bin/bash	
+#!/bin/bash
+#$ -cwd
+#$ -m e
+#$ -l mem_free=50G,h_vmem=100G,h_fsize=10G
+#$ -N ${sname}
 echo "**** Job starts ****"
 date
 
 mkdir -p ${WDIR}/${outdir}/logs
 
 # merge results
-Rscript-3.1 -e "library(derfinder); load('/dcs01/lieber/ajaffe/Brain/derRuns/derfinderExample/derGenomicState/GenomicState.Hsapiens.UCSC.hg19.knownGene.Rdata'); mergeResults(prefix='${PREFIX}', genomicState=GenomicState.Hsapiens.UCSC.hg19.knownGene)"
+module load R/3.1.x
+Rscript -e "library(derfinder); load('/dcs01/lieber/ajaffe/Brain/derRuns/derfinderExample/derGenomicState/GenomicState.Hsapiens.UCSC.hg19.knownGene.Rdata'); mergeResults(prefix='${PREFIX}', genomicState=GenomicState.Hsapiens.UCSC.hg19.knownGene$fullGenome)"
 
 # Move log files into the logs directory
 mv ${WDIR}/${sname}.* ${WDIR}/${outdir}/logs/
 
 echo "**** Job ends ****"
 date
 EOF
-call="qsub -cwd -l mem_free=50G,h_vmem=100G,h_fsize=10G -N ${sname} -m e .${sname}.sh"
+call="qsub .${sname}.sh"
 echo $call
 $call

derAnalysis/der2Models.sh

@@ -1,7 +1,7 @@
 #!/bin/sh
 
 ## Usage
-# sh der2Models.sh fetalVsInfant-v0.0.46 fetal infant
+# sh der2Models.sh run2-v0.0.42
 
 # Directories
 MAINDIR=/dcs01/lieber/ajaffe/Brain/derRuns/libd_n36
@@ -10,30 +10,33 @@ WDIR=${MAINDIR}/derAnalysis
 # Define variables
 SHORT='der2Mod-n36'
 PREFIX=$1
-GROUP1=$2
-GROUP2=$3
 
 # Construct shell files
 outdir="${PREFIX}"
 sname="${SHORT}.${PREFIX}"
 echo "Creating script ${sname}"
 cat > ${WDIR}/.${sname}.sh <<EOF
-#!/bin/bash	
+#!/bin/bash
+#$ -cwd
+#$ -m e
+#$ -l mem_free=50G,h_vmem=100G,h_fsize=10G
+#$ -N ${sname}
 echo "**** Job starts ****"
 date
 
 mkdir -p ${WDIR}/${outdir}/logs
 
 # merge results
 cd ${WDIR}/${outdir}/
-Rscript-3.1 ${WDIR}/derfinder2-models.R -r "$GROUP1" -c "$GROUP2"
+module load R/3.1.x
+Rscript ${WDIR}/derfinder2-models.R
 
 # Move log files into the logs directory
 mv ${WDIR}/${sname}.* ${WDIR}/${outdir}/logs/
 
 echo "**** Job ends ****"
 date
 EOF
-call="qsub -cwd -l mem_free=50G,h_vmem=100G,h_fsize=10G -N ${sname} -m e .${sname}.sh"
+call="qsub .${sname}.sh"
 echo $call
-$call
+$call

derAnalysis/der2Report.sh

@@ -1,7 +1,9 @@
 #!/bin/sh
 
 ## Usage
-# sh der2Report.sh run1-v0.0.46
+# sh der2Report.sh run2-v0.0.42
+
+## regionReport available at http://www.bioconductor.org/packages/release/bioc/html/regionReport.html
 
 # Directories
 MAINDIR=/dcs01/lieber/ajaffe/Brain/derRuns/libd_n36
@@ -16,21 +18,26 @@ outdir="${PREFIX}"
 sname="${SHORT}.${PREFIX}"
 echo "Creating script ${sname}"
 cat > ${WDIR}/.${sname}.sh <<EOF
-#!/bin/bash	
+#!/bin/bash
+#$ -cwd
+#$ -m e
+#$ -l mem_free=50G,h_vmem=100G,h_fsize=10G 
+#$ -N ${sname}
 echo "**** Job starts ****"
 date
 
 mkdir -p ${WDIR}/${outdir}/logs
 
 # merge results
-Rscript-3.1 -e "library(derfinderReport); load('${MAINDIR}/derCoverageInfo/fullCov.Rdata'); generateReport(prefix='${PREFIX}', browse=FALSE, nBestClusters=20, fullCov=fullCov, device='CairoPNG')"
+module load R/3.1.x
+Rscript -e "library(regionReport); load('${MAINDIR}/derCoverageInfo/fullCov.Rdata'); derfinderReport(prefix='${PREFIX}', browse=FALSE, nBestClusters=20, fullCov=fullCov, device='CairoPNG')"
 
 # Move log files into the logs directory
 mv ${WDIR}/${sname}.* ${WDIR}/${outdir}/logs/
 
 echo "**** Job ends ****"
 date
 EOF
-call="qsub -cwd -l mem_free=50G,h_vmem=100G,h_fsize=10G -N ${sname} -m e .${sname}.sh"
+call="qsub .${sname}.sh"
 echo $call
 $call

derAnalysis/derfinder2-analysis.R

@@ -3,7 +3,7 @@
 ## Load libraries
 library("getopt")
 
-## Available at https://github.com/lcolladotor/derfinder
+## Available at http://www.bioconductor.org/packages/release/bioc/html/derfinder.html
 library("derfinder")
 
 ## Specify parameters
@@ -64,16 +64,10 @@ load("models.Rdata")
 ## Load group information
 load("groupInfo.Rdata")
 
-## Load colsubsets used
-# load("colsubset.Rdata")
-
 ## Run the analysis
-# analyzeChr(chr=opt$chr, coverageInfo=data, models=models, colsubset=colsubset, cutoffFstat=1e-04, cutoffType="theoretical", nPermute=100, seeds=seq_len(100) * 20140408, maxClusterGap=3000, groupInfo=groupInfo, subject="hg19", mc.cores=opt$mcores, lowMemDir=paste0("chr", opt$chr, "/chunksDir"), verbose=opt$verbose)
 
 ## n36 analysis
-# analyzeChr(chr=opt$chr, coverageInfo=data, models=models, cutoffFstat=1e-08, cutoffType="theoretical", nPermute=1000, seeds=seq_len(1000), maxClusterGap=3000, groupInfo=groupInfo, subject="hg19", mc.cores=opt$mcores, lowMemDir=paste0("chr", opt$chr, "/chunksDir"), verbose=opt$verbose)
-## without lowMemDir
-analyzeChr(chr=opt$chr, coverageInfo=data, models=models, cutoffFstat=1e-08, cutoffType="theoretical", nPermute=1000, seeds=seq_len(1000), maxClusterGap=3000, groupInfo=groupInfo, subject="hg19", mc.cores=opt$mcores, verbose=opt$verbose)
+analyzeChr(chr=opt$chr, coverageInfo=data, models=models, cutoffFstat=1e-08, cutoffType="theoretical", nPermute=1000, seeds=seq_len(1000), maxClusterGap=3000, groupInfo=groupInfo, subject="hg19", mc.cores=opt$mcores, lowMemDir=file.path(tempdir(), paste0("chr", opt$chr), "chunksDir"), verbose=opt$verbose)
 
 ## Done
 if(opt$verbose) {

derAnalysis/derfinder2-models.R

@@ -1,40 +1,8 @@
 ## Calculate the library adjustments and build the models
 
-library("getopt")
-
-## Available at https://github.com/lcolladotor/derfinder
+## Available at http://www.bioconductor.org/packages/release/bioc/html/derfinder.html
 library("derfinder")
 
-## Specify parameters
-spec <- matrix(c(
-	'reference', 'r', 1, "character", "group 1 to use in the comparison",
-	'comparison', 'c', 1, "character", "group 2 to use in the comparison",
-	'help' , 'h', 0, "logical", "Display help"
-), byrow=TRUE, ncol=5)
-opt <- getopt(spec)
-
-## Testing the script
-test <- FALSE
-if(test) {
-	## Speficy it using an interactive R session and testing
-	test <- TRUE
-}
-
-## Test values
-if(test){
-	opt <- NULL
-	opt$group1 <- "(-1,0]"
-	opt$group2 <- "(0,1]"
-}
-
-## if help was asked for print a friendly message
-## and exit with a non-zero error code
-if (!is.null(opt$help)) {
-	cat(getopt(spec, usage=TRUE))
-	q(status=1)
-}
-
-
 ## Load the coverage information
 load("../../derCoverageInfo/filteredCov.Rdata")
 
@@ -51,20 +19,8 @@ info <- info[complete.cases(info),]
 info$dir <- paste0("R", info$RNANum)
 match <- sapply(dirs, function(x) { which(info$dir == x)})
 info <- info[match, ]
-## Set the first group as the reference
-twogroups <- c(opt$reference, opt$comparison)
-cases <- c("(-1,0]", "(0,1]", "(1,10]", "(10,20]", "(20,50]", "(50,100]")
-names(cases) <- c("fetal", "infant", "child", "teen", "adult", "elderly")
-levels <- cases[match(twogroups, names(cases))]
-
-group <- factor(info$ageGroup, levels=levels)
-
-## Define colsubset
-colsubset <- which(!is.na(group))
-save(colsubset, file="colsubset.Rdata")
-
-## Update the group labels
-group <- group[!is.na(group)]
+## Set the control group as the reference
+group <- relevel(info$ageGroup, "(0,1]")
 
 ## Determine sample size adjustments
 if(file.exists("sampleDepths.Rdata")) {
@@ -77,7 +33,7 @@ if(file.exists("sampleDepths.Rdata")) {
 		load("../../derCoverageInfo/fullCov.Rdata")
 
 		## Collapse
-		collapsedFull <- collapseFullCoverage(fullCov, colsubset=colsubset, save=TRUE)
+		collapsedFull <- collapseFullCoverage(fullCov, save=TRUE)
 	}
 
 	## Get the adjustments

derAnalysis/fix-colnames-fullRegions-2014-03-29.R

@@ -15,3 +15,5 @@ colnames(values(fullRegions))[i] <- paste("log2FoldChange", levels(groupInfo)[2:
 colnames(values(fullRegions))
 
 save(fullRegions, file="fullRegions.Rdata")
+
+## Note: this might no longer be needed with the BioC version of derfinder, but we'll leave it here just in case.

derCoverageInfo/derCoverageInfo.sh

@@ -6,31 +6,36 @@ WDIR=${MAINDIR}/libd_n36/derCoverageInfo
 DATADIR=${MAINDIR}/n36
 
 # Define variables
-SHORT='derCovInf'
+SHORT='derCovInf-n36'
 
 # Construct shell files
 for chrnum in 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y
 do
 	chr="chr${chrnum}"
 	echo "Creating script for chromosome ${chr}"
 	cat > ${WDIR}/.${SHORT}.${chr}.sh <<EOF
-#!/bin/bash	
+#!/bin/bash
+#$ -cwd
+#$ -m e
+#$ -l mem_free=50G,h_vmem=100G,h_fsize=30G
+#$ -N ${SHORT}.${chr}
 echo "**** Job starts ****"
 date
 
 # Make logs directory
 mkdir -p ${WDIR}/logs
 
-# run makeDF()
-R-devel -e "library(derfinder2); dirs <- makeBamList(datadir='${DATADIR}', samplepatt='bam$', bamterm=NULL); names(dirs) <- gsub('_accepted_hits.bam', '', names(dirs)); loadCoverage(dirs=dirs, chr='${chr}', cutoff=5, output='auto', verbose=TRUE); proc.time(); sessionInfo()"
+# run loadCoverage()
+module load R/3.1.x
+R -e "library(derfinder); files <- rawFiles(datadir='${DATADIR}', samplepatt='bam$', bamterm=NULL); names(files) <- gsub('_accepted_hits.bam', '', names(files)); loadCoverage(files=files, chr='${chr}', cutoff=5, output='auto', verbose=TRUE); proc.time(); sessionInfo()"
 
 ## Move log files into the logs directory
 mv ${SHORT}.${chr}.* ${WDIR}/logs/
 
 echo "**** Job ends ****"
 date
 EOF
-	call="qsub -cwd -l jabba,mem_free=50G,h_vmem=100G,h_fsize=30G -N ${SHORT}.${chr} -m e ${WDIR}/.${SHORT}.${chr}.sh"
+	call="qsub ${WDIR}/.${SHORT}.${chr}.sh"
 	echo $call
 	$call
 done

derCoverageInfo/mergeCov.R

@@ -1,17 +1,18 @@
 ## Merge the coverage data
+## Note that you would now use the fullCoverage(cutoff = 5) function which did not exist when this script was made.
 
-library(IRanges)
+library('IRanges')
 
 ## setup
 chrs <- c(1:22, "X", "Y")
-covList <- vector("list", 24)
-names(covList) <- chrs
+filteredCov <- vector("list", 24)
+names(filteredCov) <- chrs
 
 ## Actual processing
 for(chr in chrs) {
 	load(paste0("chr", chr, "CovInfo.Rdata"))
 	eval(parse(text=paste0("data <- ", "chr", chr, "CovInfo")))
-	covList[[chr]] <- data
+	filteredCov[[chr]] <- data
 }
 
-save(covList, file="covList.Rdata")
+save(filteredCov, file="filteredCov.Rdata")

fullCoverage/fullCoverage.sh

@@ -6,31 +6,36 @@ WDIR=${MAINDIR}/libd_n36/fullCoverage
 DATADIR=${MAINDIR}/n36
 
 # Define variables
-SHORT='fullCov'
+SHORT='fullCov-n36'
 
 # Construct shell files
 for chrnum in 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y
 do
 	chr="chr${chrnum}"
 	echo "Creating script for chromosome ${chr}"
 	cat > ${WDIR}/.${SHORT}.${chr}.sh <<EOF
-#!/bin/bash	
+#!/bin/bash
+#$ -cwd
+#$ -m e
+#$ -l mem_free=50G,h_vmem=100G,h_fsize=30G
+#$ -N ${SHORT}.${chr}
 echo "**** Job starts ****"
 date
 
 # Make logs directory
 mkdir -p ${WDIR}/logs
 
-# run makeDF()
-R-devel -e "library(derfinder2); dirs <- makeBamList(datadir='${DATADIR}', samplepatt='bam$', bamterm=NULL); names(dirs) <- gsub('_accepted_hits.bam', '', names(dirs)); loadCoverage(dirs=dirs, chr='${chr}', cutoff=NULL, output='auto', verbose=TRUE); proc.time(); sessionInfo()"
+# run loadCoverage(cutoff = NULL)
+module load R/3.1.x
+R -e "library(derfinder); files <- rawFiles(datadir='${DATADIR}', samplepatt='bam$', bamterm=NULL); names(files) <- gsub('_accepted_hits.bam', '', names(files)); loadCoverage(files=files, chr='${chr}', cutoff=NULL, output='auto', verbose=TRUE); proc.time(); sessionInfo()"
 
 ## Move log files into the logs directory
 mv ${SHORT}.${chr}.* ${WDIR}/logs/
 
 echo "**** Job ends ****"
 date
 EOF
-	call="qsub -cwd -l jabba,mem_free=50G,h_vmem=100G,h_fsize=30G -N ${SHORT}.${chr} -m e ${WDIR}/.${SHORT}.${chr}.sh"
+	call="qsub ${WDIR}/.${SHORT}.${chr}.sh"
 	echo $call
 	$call
 done

fullCoverage/mergeFull.R

@@ -1,6 +1,7 @@
 ## Merge the coverage data
+## Note that you would now use the fullCoverage() function which did not exist when this script was made.
 
-library(IRanges)
+library('IRanges')
 
 ## setup
 chrs <- c(1:22, "X", "Y")
@@ -14,4 +15,4 @@ for(chr in chrs) {
 	fullCov[[chr]] <- data$coverage
 }
 
-save(fullCov, file="fullCov.Rdata")
+save(fullCov, file="../derCoverageInfo/fullCov.Rdata")