dge_utils.sh

## docs
## http://blog.joncairns.com/2013/08/what-you-need-to-know-about-bash-functions/

## load lsf helpers
source <(curl https://raw.githubusercontent.com/holgerbrandl/datautils/v1.14/bash/lsf_utils.sh 2>&1 2>/dev/null)

## enable snippet spinning
source <(curl https://raw.githubusercontent.com/holgerbrandl/datautils/v1.14/R/spinr/spin_utils.sh 2>&1 2>/dev/null)


## define common binaries
## todo tweak this to work on bioinfo as well
export PATH=/projects/bioinfo/holger/bin/bowtie2-2.2.5:$PATH
export PATH=/projects/bioinfo/holger/bin/tophat-2.0.13.Linux_x86_64:$PATH
export PATH=/home/brandl/bin/cufflinks-2.2.1.Linux_x86_64:$PATH
export PATH=/sw/apps/python/current/bin:$PATH
export PATH=/home/brandl/bin/deepTools/bin:$PATH
export PATH=/projects/bioinfo/holger/bin/FastQC_0.11.2:$PATH
export PATH=/projects/bioinfo/holger/bin/bedtools-2.23.0/bin/:$PATH
export PATH=/home/brandl/bin/spinr:$PATH
export PATH=/home/brandl/bin/subread-1.4.6-p3-Linux-x86_64/bin:$PATH

#export PATH=/home/brandl/bin/STAR/STAR-STAR_2.4.1d/source:$PATH
export PATH=/home/brandl/bin/STAR/STAR_2.4.2a/source:$PATH



# which tophat;  which bowtie2; which cuffdiff


## 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
    

    mysub "fastqc__$(basename $fastqFile)" "fastqc -j /sw/bin/java -o $outputDir -f fastq $fastqFile" -q medium  | joblist .fastqc_jobs
done


wait4jobs .fastqc_jobs


spin.R ${NGS_TOOLS}/dge_workflow/fastqc_summary.R $outputDir

mailme "$project: fastqc done in $(pwd)"

}
export -f dge_fastqc


dge_cutadapt(){

export Ill_ADAPTERS=/projects/bioinfo/common/cutadapt_patterns.fasta

for fastqFile in $* ; do
    # DEBUG fastqFile=/projects/bioinfo/holger/projects/helin/mouse/treps_pooled/mouse_liver_polar_stage1_rep4.fastq.gz
    caFastq=$(basename $fastqFile .fastq.gz)_ca.fastq.gz
    echo "cutadapting $caFastq into $caFastq"

    #todo use a more specific trimming model (trim just correct part on each side without using reverse complements
    mysub "${project}__ca__$(basename $fastqFile .fastq.gz)" "cutadapt -b file:$Ill_ADAPTERS -m 20 -q 25 -o $caFastq $fastqFile"  -q long  | joblist .cajobs
done

wait4jobs .cajobs

spin.R ${NGS_TOOLS}/dge_workflow/cutadapt_summary.R .



## todo do a small report here about what has been trimmed away and why

}
export -f dge_cutadapt


#http://wiki.bash-hackers.org/howto/getopts_tutorial
dge_tophat_se(){

# http://stackoverflow.com/questions/18414054/bash-getopts-reading-optarg-for-optional-flags
#echo $*

while getopts "i:" curopt; do
    case $curopt in
    i) IGENOME="$OPTARG";
    esac
done
shift $(($OPTIND - 1))


local fastqFiles=$*

# IGENOME=/projects/bioinfo/igenomes/Mus_musculus/Ensembl/GRCm38

if [ -z "$IGENOME" ] || [ -z "$fastqFiles" ];
     then echo "Usage: dge_tophat_se -i <path to igenome> [<fastq.gz file>]+" >&2 ; return;
fi



export bowtie_gindex="$IGENOME/Sequence/Bowtie2Index/genome"
export gtfFile="$IGENOME/Annotation/Genes/genes.gtf"
#head $gtfFile

if [ ! -f $gtfFile ]; then
    >&2 echo "gtf '$gtfFile' does not exis"; return;
fi

if [ -z "$(which tophat)" ]; then
    >&2 echo "no tomcat binary in PATH"; return;
fi


echo "running tophat using igenome '$IGENOME' for the following files"
ll $fastqFiles

#fastqFiles=$(ls $baseDir/treps_pooled/*fastq.gz)

for fastqFile in $fastqFiles ; do
    echo "submitting tophat job for $fastqFile"

    # DEBUG fastqFile=/projects/bioinfo/holger/projects/helin/mouse/trimmed/mouse_big_cyst_rep4_ca.fastq.gz
    fastqBaseName=$(basename ${fastqFile%%.fastq.gz})
    outputdir=$fastqBaseName

    ## uniquely mapping reads only:   http:/seqanswers.com/forums/showthread.php?s=93da11109ae12a773a128a637d69defe&t=3464
    ###     tophat -p6  -G $gtfFile -g1 -o test $bowtie_gindex $fastqFile
    mysub "${project}__tophat__${fastqBaseName}" "
    tophat -p6  -G $gtfFile -g1 -o $outputdir $bowtie_gindex $fastqFile

    mv $outputdir/accepted_hits.bam $outputdir/$(basename $outputdir).bam
    samtools index $outputdir/$(basename $outputdir).bam
    " -n 5 -R span[hosts=1] -q medium | joblist .tophatjobs
done

wait4jobs .tophatjobs



## see https://github.com/fidelram/deepTools/wiki/QC#bamCorrelate
dge_bam_correlate . &

## create tophat mapping report
spin.R ${NGS_TOOLS}/dge_workflow/tophat_qc.R .

mailme "$project: tophat done in $(pwd)"

}
export -f dge_tophat_se


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

local bamDir=$1

if [ $# -e 1 ]; then
    bamFiles=$(find $bamDir | grep ".bam$" | grep -v "unmapped" | sort)
else
    bamFiles=$*
fi

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 $bamLabels


## see how well bam files correlate using untrimmed data
bcCmd="bamCorrelate bins --bamfiles $(echo $bamFiles | xargs echo) --labels $bamLabels --plotFile='bc.pdf' --outFileCorMatrix='bc.txt' --numberOfProcessors=6 --corMethod spearman --zMin 0.5 --zMax 1"
echo $bcCmd
#mysub "${project}__bamcorrelate" "$bcCmd"  -q long -n 4 -R span[hosts=1] | blockScript .bamcorrelate
#mailme "$project: bamcorrelate done in $(pwd)"

}
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 $bamFile to bigwig format"
    mysub "${project}__bw__${sample}" "genomeCoverageBed -split -bg -ibam $bamFile  -g ${genomeFai} |  wigToBigWig -clip stdin ${genomeFai} ${sample}.bw" -q short | joblist .bigwig
done

wait4jobs .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 bsub 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

## tester
#dge_merge_treps $baseDir/mapped_trim/ "Ctrl_0703,Ctrl_1029,Ctrl_1103,Insm1_0703,Insm1_1029,Insm1_1103"



#dge_cd(){
#( ## required to work around docopt sys.exit
#usage='
#Use cuffdiff2 to performa a differntial expression analysis
#Usage: dge_cuffdiff [--exclude=<regex>] <gtfFile> <bamDir> <labels>
#
#Options:
#--exclude=<regex>   Exclude patterns (grep regex matching bam file paths to be excluded)
#'
#echo "$usage" | ~/bin/docopts/docopts -h -  : $*
#eval "$(echo "$usage" | ~/bin/docopts/docopts -h -  : $*)"
#
#)
#}


#dge_cd $gtfFilePC $baseDir/mapped $labels
#dge_cd --help


dge_cuffdiff(){

local gtfFile=$1
local bamDir=$2
local labels=$3

if [ $# -ne 3 ]; then
    echo "Usage: dge_cuffdiff <gtf_file>  <bam_directory> <labels>" >&2 ; return;
fi

if [ -z "$(which cuffdiff)" ]; then
    >&2 echo "no cuffdiff binary in PATH"; return;
fi

if [ ! -f $gtfFile ]; then
    >&2 echo "gtf '$gtfFile' does not exis"; return;
fi

## collect all bam-files and group them
allBams=$(find $bamDir | grep ".bam$" | grep -v "unmapped" | sort)

if [ -n "$EXCLUDE_BAMS_PATTERN" ]; then
    echo "excluding bams with $EXCLUDE_BAMS_PATTERN"
    allBams=$(echo "$allBams" | grep -v "$EXCLUDE_BAMS_PATTERN")
fi

## todo use optional argument here --> default "unmapped" --> allow user to add others
#allBams=$(find $bamDir | grep ".bam$" | grep -v "unmapped" | grep -v "1029" | sort)
#allBams=$(find $bamDir | grep ".bam$" | grep -v "unmapped" | grep -v "Ctrl_1029" | sort)

bamsSplit=""
for label in $(echo $labels | tr ", " " "); do
    # DEBUG label="liver"; label="cyst"
    echo $label

    ## grep the bam names excluding the path
    echo  "$allBams" | xargs -n1 basename | grep $label > ${label}.cuff_bamlist
    labelBams=$(echo "$allBams" | grep -Ff ${label}.cuff_bamlist | xargs echo -n | tr ' ' ',')
    # echo "$allBams" | grep -Ff ${label}.bamlist | xargs -n1 echo

    bamsSplit+=$labelBams" "
done
#echo $bamsSplit

## make sure (again that all files are there
echo $gtfFile $bamsSplit | tr "," " "  | xargs ls -la


#mysub ${project}_cuffdiff "cuffdiff -L aRGm,aRGp,aRG,bRG,IPC,N -o . -p 8 mm10_ensGene_ccds.gtf $amBams $apBams $aBams $bBams $cBams $dBams  2>&1 | tee cuffdiff.log" -q long -n 8 -R span[hosts=1]   | blockScript

cdCmd="cuffdiff -L $labels -o . -p 10 $gtfFile $bamsSplit"
echo "cuffdiff cmd is: $cdCmd"
#eval $cdCmd
mysub "${project}__cuffdiff" "$cdCmd"  -q long -n 4 -R span[hosts=1] | blockScript


## create a cuffdiff db using cummerbund in a temporary directory to avoid locking problems
tmpDbDir=$(mktemp -d)
cp -r . $tmpDbDir

## todo remove this hack
genome=$(echo $gtfFile | cut -f8 -d'/' | tr '[:upper:]' '[:lower:]'); echo "genome is $genome"

## make sure to use temp-r to avoid file locking problems
#export R_LIBS=/tmp/r_index

echo '
require(cummeRbund)
dbDir=commandArgs(T)[1]
gtfFile=commandArgs(T)[2]
genome=commandArgs(T)[3]
## note without providing the gtf the db is much smaller
readCufflinks(dir=dbDir, rebuild=T, gtf=gtfFile, genome=genome)
' | R -q --no-save --no-restore --args "$tmpDbDir" "$gtfFile" "$genome"

if [ ! -f $tmpDbDir/cuffData.db ]; then
    >&2 echo "cummerbund failed to create cuffidff sqlite db"; return;
fi

cp $tmpDbDir/cuffData.db .
rm -rf $tmpDbDir ## because it's no longer needed

mailme "$project: cuffdiff done in $(pwd)"

}
export -f dge_cuffdiff


# 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 ${igenome}/Sequence/Chromosomes/*.fa --runThreadN 10
    mysub "${project}_star_index" "$cmd" -n 5 -R span[hosts=1] -q medium  | blockScript

    ## prevent further modification
    chmod -w $(dirname ${star_index})

    mailme "created star index for $igenome"
}
export -f dge_create_star_index


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("gene_id", "num_alignments") %>%
        filter(!str_detect(gene_id, "^N_")) %>%
        mutate(sample=trim_ext(countFile, ".ReadsPerGene.out.tab"))
}, .progress="text")

countMatrix <- spread(exprCounts, sample, num_alignments)

write.delim(countMatrix, "star_count_matrix.txt")
' | R --vanilla -q

}
export -f dge_star_counts2matrix