Is this the correct order of steps in GATK germline cohort variant calling (VQSR workflow)?
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16 hours ago
Ramnaresh • 0

I’m performing germline variant calling using GATK (gVCF-based workflow) and would like to confirm whether the order of my steps is correct. What I have done so far:

CombineGVCFs            ==> cohort.g.vcf.gz
GenotypeGVCFs           ==> output.vcf.gz
VariantFiltration (ExcessHet) ==> cohort_excesshet.vcf.gz
bcftools norm -m-any --check-ref -w -f "$reference" cohort_excesshet.vcf.gz -o cohort.nm.vcf.gz
MakeSitesOnlyVcf        ==> cohort.sitesonly.vcf.gz
VQSR

I’m trying to generate a high-quality cohort VCF and will later analyze per-patient variants. Is this the correct order of steps? Should normalization (bcftools norm) or VariantFiltration be performed before or after VQSR?

System:
GATK 4.6.2.0
Reference: GRCh38
~140 samples (gVCFs)
16 GB RAM

Any suggestions or corrections are appreciated!

Thanks in advance.

gVCF VCF GATK VQSR • 70 views
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