Hi, I just finished running HaplotypeCaller version 3.3-0 separately on 6 exome samples with the new best practices.
java -Xmx8g -jar GenomeAnalysisTK.jar -T HaplotypeCaller -R hg19/hg19_Ordered.fa -I K87/HG19_Analysis/K87-929_final.recalibrated_final.bam --dbsnp dbsnp_138_hg19_Ordered.vcf --pair_hmm_implementation VECTOR_LOGLESS_CACHING -ERC GVCF -variant_index_type LINEAR -variant_index_parameter 128000 --output_mode EMIT_VARIANTS_ONLY -gt_mode DISCOVERY --pcr_indel_model CONSERVATIVE -o ./Haplotypes_929.vcf
Many Variant sites are called as homozygous alt (1/1), but none of these sites that are processed to infer haplotype are called as homozygous alt in their PGT field, they are all called as hets, PGT=0|1. for example:
GT:AD:DP:GQ:PGT:PID:PL:SB 1/1:0,29,0:29:93:0|1:121483392_C_G:1331,93,0,1331,93,1331:0,0,13,16
The allelic depths agree with the phased genotype but out of all 6 exomes processed, not a single 1/1 is also phased as 1|1.
I checked all output vcfs with a simple grep combo: grep 'PGT' Haplotypes_929.vcf | grep '1/1' - | grep '0|1' - | wc -l = 19046 grep 'PGT' Haplotypes_929.vcf | grep '1/1' - | grep '1|0' - | wc -l = 79 grep 'PGT' Haplotypes_929.vcf | grep '1/1' - | grep '1|1' - | wc -l = 0
This seemed odd, but I continued with GenotyeGVCF:
java -Xmx32g -jar GenomeAnalysisTK.jar -T GenotypeGVCFs -R hg19/hg19_Ordered.fa -V Haplotypes_450.vcf -V Haplotypes_452.vcf -V Haplotypes_925.vcf -V Haplotypes_926.vcf -V Haplotypes_927.vcf -V Haplotypes_929.vcf -D dbsnp_138_hg19_Ordered.vcf -ped K87/HG19_Analysis/K87_6.ped -o Haplotypes_K87_GVCFs.vcf
I'm looking at the output vcf as it's being generated and now there are homozygous alt calls but they conflict with the associated Allelic Depths:
.... GT:AD:DP:GQ:PGT:PID:PL .... 1/1:0,29:29:85:1|1:33957151_G_T:948,85,0 .....
Full Line: chr1 33957152 rs4403594 T G 3166.96 . AC=12;AF=1.00;AN=12;DB;DP=99;FS=0.000;GQ_MEAN=48.50;GQ_STDDEV=27.55;MLEAC=12;MLEAF=1.00;MQ=39.65;MQ0=0;NCC=0;QD=32.32;SOR=0.693 GT:AD:DP:GQ:PGT:PID:PL 1/1:0,9:9:27:.:.:330,27,0 1/1:0,5:5:15:.:.:141,15,0 1/1:0,29:29:85:1|1:33957151_G_T:948,85,0 1/1:0,20:20:60:.:.:722,60,0 1/1:0,24:24:71:.:.:685,71,0 1/1:0,11:11:33:.:.:366,33,0
Can you help me interpret what seems to me as conflicting results?
Cheers,
Patrick