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variant_germline
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7cddb73f
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7cddb73f
authored
8 years ago
by
Brandi Cantarel
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bugs with gatk
parent
bb09df55
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workflow/main.nf
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7cddb73f
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@@ -10,7 +10,7 @@ params.capture="$params.genome/PanCancerTargetRegions.hg38.bed"
dbsnp="$params.genome/dbSnp.vcf.gz"
indel="$params.genome/GoldIndels.vcf.gz"
design_file = file(params.design)
btoolsgenome = file("$params.genome/genomefile.txt")
fastqs=file(params.fastqs)
knownindel=file(indel)
gatkref=file("$params.genome/genome.fa")
...
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@@ -175,7 +175,7 @@ process seqqc {
sambamba flagstat -t 30 ${sbam} > ${pair_id}.flagstat.txt
sambamba view -t 30 -f bam -F "mapping_quality >= 1" -L ${capture_bed} -o ${pair_id}.ontarget.bam ${sbam}
sambamba flagstat -t 30 ${pair_id}.ontarget.bam > ${pair_id}.ontarget.flagstat.txt
bedtools coverage -sorted -hist -g ${
btoolsgenome}
-b ${pair_id}.ontarget.bam -a ${capture_bed} | grep ^all > ${pair_id}.genomecov.txt
bedtools coverage -sorted -hist -g ${
index_path}/genomefile.txt
-b ${pair_id}.ontarget.bam -a ${capture_bed} | grep ^all > ${pair_id}.genomecov.txt
perl $baseDir/scripts/subset_bam.pl ${pair_id}.ontarget.bam ${pair_id}.ontarget.flagstat.txt
java -Xmx8g -jar \$PICARD/picard.jar EstimateLibraryComplexity INPUT=${pair_id}.subset50M.bam OUTPUT=${pair_id}.libcomplex.txt
"""
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@@ -250,11 +250,11 @@ process gatkgvcf {
input:
set pair_id,file(gbam),file(gidx) from gatkbam
output:
file("${pair_id}.gatk.gvcf") into gatkgvcf
file("${pair_id}.gatk.g
.
vcf") into gatkgvcf
script:
"""
module load gatk/3.5 bedtools/2.25.0 snpeff/4.2 vcftools/0.1.11
java -Xmx10g -jar \$GATK_JAR -R ${gatkref} -D ${dbsnp} -T HaplotypeCaller -stand_call_conf 30 -stand_emit_conf 10.0 -A FisherStrand -A QualByDepth -A VariantType -A DepthPerAlleleBySample -A HaplotypeScore -A AlleleBalance -variant_index_type LINEAR -variant_index_parameter 128000 --emitRefConfidence GVCF -I ${gbam} -o ${pair_id}.g.vcf -nct 2
java -Xmx10g -jar \$GATK_JAR -R ${gatkref} -D ${dbsnp} -T HaplotypeCaller -stand_call_conf 30 -stand_emit_conf 10.0 -A FisherStrand -A QualByDepth -A VariantType -A DepthPerAlleleBySample -A HaplotypeScore -A AlleleBalance -variant_index_type LINEAR -variant_index_parameter 128000 --emitRefConfidence GVCF -I ${gbam} -o ${pair_id}.
gatk.
g.vcf -nct 2
"""
}
process gatk {
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@@ -290,7 +290,7 @@ process mpileup {
"""
module load samtools/intel/1.3 bedtools/2.25.0 bcftools/intel/1.3 snpeff/4.2 vcftools/0.1.11
ls *.bam > final.bam.list
cut -f 1 ${index_path}/genomefile.txt | xargs -I {} -n 1 -P 32 sh -c "samtools mpileup -t 'AD,ADF,ADR,INFO/AD,SP' -ug -Q20 -C50 -f ${index_path}/${index_name}.fa -b final.bam.list -r {} | bcftools call --format-fields gq,gp -vmO z -o final.sam.{}.vcf.gz"
cut -f 1 ${index_path}/genomefile.
chr.
txt | xargs -I {} -n 1 -P 32 sh -c "samtools mpileup -t 'AD,ADF,ADR,INFO/AD,SP' -ug -Q20 -C50 -f ${index_path}/${index_name}.fa -b final.bam.list -r {} | bcftools call --format-fields gq,gp -vmO z -o final.sam.{}.vcf.gz"
vcf-concat final.sam.*.vcf.gz | vcf-sort |vcf-annotate -n --fill-type | java -jar \$SNPEFF_HOME/SnpSift.jar filter '((QUAL >= 10) & (MQ >= 20) & (DP >= 10))' |bedtools intersect -header -a stdin -b ${capture_bed} |bgzip > final.sampanel.vcf.gz
"""
}
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