diff --git a/alignment/bamqc.sh b/alignment/bamqc.sh index 44ba8cdf53cef29452af150fd0ea4c5dad4ae50b..566eea451e0332e5868311645fcdd198ac22acd3 100644 --- a/alignment/bamqc.sh +++ b/alignment/bamqc.sh @@ -48,10 +48,10 @@ if [[ $nuctype == 'dna' ]]; then samtools flagstat ${pair_id}.ontarget.bam > ${pair_id}.ontarget.flagstat.txt samtools view -b -q 1 ${pair_id}.ontarget.bam | bedtools coverage -sorted -hist -g ${index_path}/genomefile.txt -b stdin -a ${bed} > ${pair_id}.mapqualcov.txt samtools view -b -F 1024 ${pair_id}.ontarget.bam | bedtools coverage -sorted -g ${index_path}/genomefile.txt -a ${bed} -b stdin -hist | grep ^all > ${pair_id}.dedupcov.txt - java -Xmx32g -jar $PICARD/picard.jar CollectAlignmentSummaryMetrics R=${reffa} I=${pair_id}.ontarget.bam OUTPUT=${pair_id}.alignmentsummarymetrics.txt + java -Xmx32g -jar $PICARD/picard.jar CollectAlignmentSummaryMetrics R=${index_path}/genome.fa I=${pair_id}.ontarget.bam OUTPUT=${pair_id}.alignmentsummarymetrics.txt java -Xmx64g -jar $PICARD/picard.jar EstimateLibraryComplexity I=${pair_id}.ontarget.bam OUTPUT=${pair_id}.libcomplex.txt samtools view -F 1024 ${pair_id}.ontarget.bam | awk '{sum+=$5} END { print "Mean MAPQ =",sum/NR}' > ${pair_id}.meanmap.txt - java -Xmx4g -jar $PICARD/picard.jar CollectInsertSizeMetrics INPUT=${pair_id}.bam HISTOGRAM_FILE=${pair_id}.hist.ps REFERENCE_SEQUENCE=${reffa} OUTPUT=${pair_id}.hist.txt + java -Xmx4g -jar $PICARD/picard.jar CollectInsertSizeMetrics INPUT=${sbam} HISTOGRAM_FILE=${pair_id}.hist.ps REFERENCE_SEQUENCE=${index_path}/genome.fa OUTPUT=${pair_id}.hist.txt samtools view -b -q 1 ${pair_id}.ontarget.bam | bedtools coverage -sorted -hist -g ${index_path}/genomefile.txt -b stdin -a ${bed} > ${pair_id}.mapqualcov.txt bedtools coverage -sorted -g ${index_path}/genomefile.txt -a ${bed} -b ${sbam} -hist > ${pair_id}.covhist.txt perl $baseDir/scripts/calculate_depthcov.pl ${pair_id}.covhist.txt diff --git a/alignment/dnaseqalign.sh b/alignment/dnaseqalign.sh index f0042b4e023989924f811bfd5e27c4e78e93d54e..0e60add806be8ac7c29243512146683a63dce889 100644 --- a/alignment/dnaseqalign.sh +++ b/alignment/dnaseqalign.sh @@ -42,14 +42,14 @@ baseDir="`dirname \"$0\"`" if [[ $fq1 == $fq2 ]] then - bwa mem -M -t $SLURM_CPUS_ON_NODE -R "@RG\tID:${pair_id}\tLB:tx\tPL:illumina\tPU:barcode\tSM:${pair_id}" ${index_path}/genome.fa ${fq1} > out.sam + bwa mem -M -t $SLURM_CPUS_ON_NODE -R "@RG\tID:${pair_id}\tLB:tx\tPL:illumina\tPU:barcode\tSM:${pair_id}" ${index_path}/genome.fa ${fq1} > out.sam else - bwa mem -M -t $SLURM_CPUS_ON_NODE -R "@RG\tID:${pair_id}\tLB:tx\tPL:illumina\tPU:barcode\tSM:${pair_id}" ${index_path}/genome.fa ${fq1} ${fq2} > out.sam + bwa mem -M -t $SLURM_CPUS_ON_NODE -R "@RG\tID:${pair_id}\tLB:tx\tPL:illumina\tPU:barcode\tSM:${pair_id}" ${index_path}/genome.fa ${fq1} ${fq2} > out.sam fi if [[ $umi=='umi' ]] then - k8 /cm/shared/apps/bwakit/0.7.15/bwa-postalt.js -p ${pair_id}.hla ${index_path}/genome.fa.alt out.sam | python ${baseDir}/add_umi_bam.py -s - -o output.unsort.bam + k8 /cm/shared/apps/bwakit/0.7.15/bwa-postalt.js -p ${pair_id}.hla ${index_path}/genome.fa.alt out.sam | python ${baseDir}/add_umi_sam.py -s - -o output.unsort.bam elif [[ $index_path == '/project/shared/bicf_workflow_ref/GRCh38' ]] then k8 /cm/shared/apps/bwakit/0.7.15/bwa-postalt.js -p ${pair_id}.hla ${index_path}/genome.fa.alt out.sam| samtools view -1 - > output.unsort.bam diff --git a/alignment/markdups.sh b/alignment/markdups.sh index 7bf470905630f979a065d35d6b652c0af1fd8dc3..793dc9687f9ef06efedaf47361dd0028fb4646ad 100644 --- a/alignment/markdups.sh +++ b/alignment/markdups.sh @@ -56,7 +56,7 @@ elif [ $algo == 'fgbio_umi' ] then samtools index -@ $SLURM_CPUS_ON_NODE ${sbam} source activate fgbiotools - fgbio GroupReadsByUmi -s identity -i ${sbam} -o ${pair_id}.group.bam -m 10 + fgbio GroupReadsByUmi -s identity -i ${sbam} -o ${pair_id}.group.bam -m 0 fgbio CallMolecularConsensusReads -i ${pair_id}.group.bam -p consensus -M 1 -o ${pair_id}.consensus.bam -S ':none:' source deactivate module load bwa/intel/0.7.15