## docs ## http://blog.joncairns.com/2013/08/what-you-need-to-know-about-bash-functions/ ## define common binaries if [ $(hostname) == "bioinformatics-srv1" ]; then export BIO_BIN_BASE="/local/home/brandl/bin" else export BIO_BIN_BASE="/projects/bioinfo/holger/bin" fi export PATH=${BIO_BIN_BASE}/bowtie2-2.2.5:$PATH export PATH=${BIO_BIN_BASE}/deepTools-2.2.2/bin:$PATH export PATH=${BIO_BIN_BASE}/FastQC_0.11.2:$PATH export PATH=${BIO_BIN_BASE}/bedtools2-2.25.0/bin/:$PATH export PATH=${BIO_BIN_BASE}/samtools-1.3:$PATH export PATH=${BIO_BIN_BASE}/STAR-2.5.1b/source:$PATH #export PATH=/home/brandl/bin/subread-1.4.6-p3-Linux-x86_64/bin:$PATH ## add cluster job manager export PATH=/projects/bioinfo/tools/joblist_v0.6:$PATH ## makd sure that rend.R is present source <(curl https://raw.githubusercontent.com/holgerbrandl/datautils/v1.23/R/rendr/rendr_utils.sh 2>&1 2>/dev/null) ## backward compatibility wrappers for old shell cluster utils #joblist(){ # jl add $* #} #export -f joblist # # #wait4jobs(){ # jl wait $* #} #export -f wait4jobs # which tophat; which bowtie2; which cuffdiff mcdir(){ if [ ! -d "$1" ]; then mkdir "$1"; fi; cd "$1"; } export -f mcdir mailme(){ echo "Subject:"$1 "$2" | sendmail -v $(whoami)@mpi-cbg.de > /dev/null ; } export -f mailme ziprm(){ if [ $# -lt 2 ]; then echo "Usage: ziprm <tarbasename> [<file>]+"; return; fi tarName=$(date +'%y%m%d')_"$1".tar.gz; shift tar czf $tarName $@; rm $@; } export -f ziprm ## no longer needed because packages are no kept in home #export R_LIBS=/tmp/r_index ## export to make sure that packages are load from local repository, otherwise sqlite won't work ## create fastq report for all fastq and fastq.gz files in the current directory dge_fastqc(){ while getopts "o:" curopt; do case $curopt in o) outputDir=$OPTARG; esac done shift $(($OPTIND - 1)) local fastqFiles=$* #if [ -z "$fastqFiles" ]; then if [ $# -lt 1 ]; then echo "Usage: dge_fastqc [-o <output_directory>] [<fastq.gz file>]+" >&2 ; return; fi ## use current directory if not specified if [ -z "$outputDir" ]; then outputDir="fastqc" fi if [ ! -d "$outputDir" ]; then echo "creating output directory '$outputDir'" mkdir $outputDir fi for fastqFile in $fastqFiles ; do echo "fastqcing $fastqFile" if [ ! -f $fastqFile ]; then continue; fi jl submit -j .fastqc_jobs -q medium -n "fastqc__$(basename $fastqFile)" "fastqc -j ${JAVA_HOME}/bin/java -o $outputDir -f fastq $fastqFile" done jl wait --report .fastqc_jobs rend.R ${NGS_TOOLS}/misc/fastqc_summary.R $outputDir mailme "$project: fastqc done in $(pwd)" } export -f dge_fastqc dge_bam_correlate(){ if [ $# -eq 0 ]; then echo "Usage: dge_bam_correlate <bam_directory>" >&2 ; echo "Usage: dge_bam_correlate <bam_files...>" >&2 ; return; fi if [ $# -eq 1 ]; then bamFiles=$(find $1 | grep ".bam$" | grep -v "unmapped" | sort) else bamFiles=$* fi local bamLabels=$(echo "$bamFiles" | xargs -n1 basename | sed 's!.*/!!' | sed 's/_mmf.bam//g' | sed 's/_ca.bam//g' | sed 's/.bam//g' | xargs echo); echo "Used bam labels are: $bamLabels" ## see how well bam files correlate using untrimmed data # http://deeptools.readthedocs.org/en/latest/content/tools/multiBamSummary.html # http://deeptools.readthedocs.org/en/latest/content/tools/plotCorrelation.html?highlight=plotfile bcCmd=" ## todo add python2 ~/.local/bin/ or fix setup multiBamSummary bins --bamfiles $(echo $bamFiles | xargs echo) --labels $bamLabels -out bam_cor_matrix.txt --numberOfProcessors 4 plotCorrelation --corData bam_cor_matrix.txt --plotFile bc.pdf --corMethod spearman --zMin 0.5 --zMax 1 -p heatmap " #echo "cmd is $bcCmd" #mysub "${project}__bamcorrelate" "$bcCmd" -q long -n 4 -R span[hosts=1] | blockScript .bamcorrelate ## old lsf version #jl submit --wait --jl .bamcorrelate -q long -t 4 -n "${project}__bamcorrelate" "$bcCmd" jl reset .bamcorrelate jl submit --jl .bamcorrelate -w 10:00 -t 4 -n "${project}__bamcorrelate" "$bcCmd" } export -f dge_bam_correlate dge_bigwig(){ usage="Usage: dge_bigwig <genome_fai> [<bam_file>]+" if [ $# -lt 2 ]; then echo ${usage} >&2 ; return; fi #bamFiles=$(find . -name "*.bam" | grep -v unmapped | xargs echo) genomeFai=$1 bamFiles="${@:2}" if [ ! -f "${genomeFai}" ]; then echo "could not find fai index $1! ${usage}" >&2 ; return; fi ## add bigwig to PATH export PATH=~/bin:${PATH} if [ -z "$(which wigToBigWig 2>/dev/null)" ]; then echo "could not find wigToBigWig in PATH! ${usage}" >&2 ; #return; fi ## create big wig files for bamFile in $bamFiles; do sample=$(basename $bamFile .bam) echo "converting $bamFile to bigwig format" jl submit -j .bigwig -w 10:00 -n "${project}__bw__${sample}" "genomeCoverageBed -split -bg -ibam $bamFile -g ${genomeFai} | wigToBigWig -clip stdin ${genomeFai} ${sample}.bw" done jl wait --report .bigwig } export -f dge_bigwig ## http://stackoverflow.com/questions/6916856/can-bash-show-a-functions-definition #type dge_bigwig ## Merge technical replicatews dge_merge_treps(){ usage="Usage: dge_merge_treps <bam_directory> <comma_sep_biosample_spec>" if [ $# -ne 2 ]; then echo $usage >&2 ; return; fi local bam_dir=$1; # bam_dir=$baseDir/mapped_trim/ local bio_reps=$2; # bio_samples="test,lala" if [ ! -d "$bam_dir" ]; then echo "bam file directory does not exist! $usage'" >&2 ; return; fi if [ $(echo "$bio_samples" | grep "," | wc -l ) -ne 1 ]; then echo "Invalid biosample spec! $usage'" >&2 ; return; fi #for sample in aRG bRG N; do for sample in $(echo $bio_samples | tr ",", " "); do # DEBUG sample=Insm1_1103 ## todo use joblist instead here ( sampleBams=$(find $bam_dir -name '*bam' | grep -v unmapped | grep $sample) echo "merging $sample with $sampleBams" ## todo add read-groups to bam files if [ 1 -eq $(echo "$sampleBams" | wc -l) ]; then cp $sampleBams $sample.bam else samtools merge - $(echo $sampleBams | xargs echo) | samtools sort - $sample fi samtools index $sample.bam )& done } export -f dge_merge_treps # Create a star index for a given igenome dge_create_star_index(){ if [ $# -ne 1 ]; then echo "Usage: dge_create_star_index <igenome>" >&2 ; return; fi igenome=$1 if [ ! -d "$igenome" ] | [ ! -d "${igenome}/Sequence" ]; then echo "igenome directory '$igenome' does not exist" >&2 ; return; fi export star_index="${igenome}/Sequence/StarIndex" ## stop if index exists already if [ -d "$star_index" ]; then echo "Error: Index directory ${star_index} already exists." >&2 ; return; fi # chmod +w $(dirname ${star_index}) mailme "${project}: creating STAR index in ${star_index}" mkdir ${star_index} cmd="STAR --runMode genomeGenerate --genomeDir ${star_index} --genomeFastaFiles ${igenome}/Sequence/WholeGenomeFasta/genome.fa --runThreadN 10" #eval $cmd #STAR --runMode genomeGenerate --genomeDir ${star_index} --genomeFastaFiles ${igenome}/Sequence/Chromosomes/*.fa --runThreadN 10 jl submit --wait -t 5 -w 10:00 -m 30g -n "${project}_star_index" "$cmd" ## prevent further modification # chmod -w $(dirname ${star_index}) mailme "created star index for $igenome" } export -f dge_create_star_index dge_get_pc_isoforms(){ # todo write more generic version that also filtered provided gtf and/or allow for ccds filtering as well if [ $# -ne 1 ]; then echo "Usage: dge_get_pc_isoforms <hsapiens/mmusculus/other_ensembl_species_identifier>" >&2 ; return; fi echo ' require(biomaRt) require(dplyr) require(ggplot2) mart <- useDataset(paste0(commandArgs(T)[1], "_gene_ensembl"), mart = useMart("ensembl")) #mart <- useDataset("hsapiens_gene_ensembl", mart = useMart("ensembl")) #mart <- useDataset("mmusculus_gene_ensembl", mart = useMart("ENSEMBL_MART_ENSEMBL", host="www.ensembl.org")) pcTx <- getBM(attributes=c("ensembl_gene_id", "ensembl_transcript_id", "gene_biotype", "transcript_biotype"), mart=mart) %>% filter(transcript_biotype=="protein_coding") #ggplot(pcTx, aes(gene_biotype)) + geom_bar() + coord_flip() #ggplot(pcTx, aes(transcript_biotype)) + geom_bar() + coord_flip()cd #write.table(with(pcTx, data.frame(ensembl_transcript_id)), col.names=F, file="mm10_pc_tx.txt",quote=F,row.names=F) # just print results to stdout write.table(with(pcTx, data.frame(ensembl_transcript_id)), col.names=F, file=stdout(),quote=F,row.names=F) ' | Rscript --vanilla - $1 2>/dev/null } export -f dge_get_pc_isoforms dge_star_counts2matrix(){ echo ' devtools::source_url("https://raw.githubusercontent.com/holgerbrandl/datautils/v1.13/R/core_commons.R") ## STAR count file format is #column 1: gene ID #column 2: counts for unstranded RNA-seq #column 3: counts for the 1st read strand aligned with RNA (htseq-count option -s yes) exprCounts <- list.files(".", "ReadsPerGene.out.tab") %>% ldply(function(countFile){ read.delim(countFile, header=F) %>% select(V1, V2) %>% set_names("ensembl_gene_id", "num_alignments") %>% filter(!str_detect(ensembl_gene_id, "^N_")) %>% mutate(sample=trim_ext(countFile, ".ReadsPerGene.out.tab")) }, .progress="text") countMatrix <- spread(exprCounts, sample, num_alignments) write.delim(countMatrix, "star_counts_matrix.txt") ' | R --vanilla -q } export -f dge_star_counts2matrix