Vcf Tools Error

Question

[tpeters1213]

I'm working on a project that involves merging four VCF files. The first three datasets ran smoothly through vcftools' vcf-merge, but merging the final dataset produces the following error:

Uh: 4 vs 1

at Vcf.pm line 169

Vcf::throw('Vcf4_0=HASH(0xa2d9870)', 'Uh: 4 vs 1\x{a}') called at Vcf.pm line 1492

VcfReader::format_haplotype('Vcf4_0=HASH(0xa2d9870)', 'ARRAY(0xa3e65c0)', 'ARRAY(0xa412cb0)') called at /path/to/vcftools_0.1.6/perl/vcf-merge line 426

main::merge_vcf_files('HASH(0x9f54170)') called at /path/to/vcftools_0.1.6/perl/vcf-merge line 12

All files have been compressed with bgzip, indexed with tabix, and should be in proper VCF4 format.

[Zev.Kronenberg]

Where are your VCFs coming from? can you post a few lines of your VCF? My guess is that your VCFs are wonky, no fault of yours. I can probably hack VCF.pm to fix your problem.

[tpeters1213]

The first three VCF files (the ones that successfully merged together) are from the uk10k project; the last one is from 1,000 genomes.

A few lines from the uk10k datasets (only the first few patients are shown):

1 14976 rs71252251 G A,C 999 MinVQSLOD BaseQRankSum=-8.331;CSQ=ENST00000423562:WASH7P:WITHIN_NON_CODING_GENE+ENST00000430492:WASH7P:DOWNSTREAM+ENST00000438504:WASH7P:WITHIN_NON_CODING_GENE+ENST00000450305:DDX11L1:DOWNSTREAM+ENST00000456328:DDX11L1:DOWNSTREAM+ENST00000488147:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000515242:DDX11L1:DOWNSTREAM+ENST00000518655:DDX11L1:DOWNSTREAM+ENST00000537342:WASH7P:DOWNSTREAM+ENST00000538476:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000541675:WASH7P:WITHIN_NON_CODING_GENE;DP4=292802,217007,21432,16455;DP=558725;Dels=0.00;FS=633.847;HaplotypeScore=2.5910;InbreedingCoeff=-0.2474;MDV=100;MQ0=183453;MQ=19;MQRankSum=87.614;MSD=375;MSQ=99;PV0=0.001;PV1=7.6e-06;PV2=1;PV3=1;PV4=0.001,7.6e-06,1,1;QD=1.32;ReadPosRankSum=20.051;SB=0.4833;VDB=0.0434;VQSLOD=-26.1505;culprit=MQ;AN=662;AC=9,0 GT:DP:DV:GQ:PL:QP:SP 0/0:179:20:99:0,236,213,255,255,255:50:3 0/0:73:0:99:0,220,180,220,180,180:40:0 0/0:150:17:99:0,168,160,255,208,229:49:2

1 15118 rs71252250 A G 999 MinVQSLOD BaseQRankSum=119.682;CSQ=ENST00000423562:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000430492:WASH7P:DOWNSTREAM+ENST00000438504:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000450305:DDX11L1:DOWNSTREAM+ENST00000456328:DDX11L1:DOWNSTREAM+ENST00000488147:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000515242:DDX11L1:DOWNSTREAM+ENST00000518655:DDX11L1:DOWNSTREAM+ENST00000537342:WASH7P:DOWNSTREAM+ENST00000538476:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC+ENST00000541675:WASH7P:WITHIN_NON_CODING_GENE,INTRONIC;DP4=6455,94784,3011,38261;DP=149166;Dels=0.00;FS=0.000;HRun=0;HWE=0.000000;HaplotypeScore=0.4952;ICF=-0.20697;InbreedingCoeff=-0.4571;MDV=77;MQ0=96363;MQ=11;MQRankSum=14.607;MSD=148;MSQ=68;PV0=3.8e-10;PV1=1;PV2=1;PV3=1;PV4=3.8e-10,1,1,1;QD=1.58;ReadPosRankSum=-2.311;SB=0.0760;VDB=0.0421;VQSLOD=-39.5528;culprit=MQ;AN=306;AC=48 GT:DP:DV:GQ:PL:QP:SP 0/0:62:12:35:0,35,56:45:0 0/0:22:1:43:0,63,37:25:0

1 120994 . AAT A 999 EndDistBias CSQ=ENST00000466430:ENSG00000238009:UPSTREAM+ENST00000471248:ENSG00000238009:WITHI N_NON_CODING_GENE,INTRONIC+ENST00000477740:ENSG00000238009:WITHIN_NON_CODING_GENE,INTRONIC;DP4=5,34528,0,679;DP=59595;HWE=0.000029;ICF=0.10466;IND EL;IS=12,0.123711;MDV=15;MQ=32;MSD=78;MSQ=99;PV0=1;PV1=1;PV2=2.3e-70;PV3=1.9e-72;PV4=1,1,2.3e-70,1.9e-72;QD=0.0823;SB=0.0015;VDB=0.0233;AN=118;AC= 1 GT:DP:DV:GQ:PL:QP:SP ./.:7:0:35:0,21,53:28:0 0/0:30:0:99:0,90,165:40:0 ./.:0:0:0:0,0,0:.:0 ./.:5:0:29:0,15,12:13:0 0/0:18:0:68:0,54 ,158:39:0 ./.:4:0:26:0,12,66:30:0 ./.:6:0:32:0,18,72:31:0 0/0:21:0:77:0,63,160:39:0 0/0:16:0:62:0,48,154:39:0 ./.:2:0:20:0,6,45: 27:0 ./.:1:0:17:0,3,23:20:0 ./.:14:0:56:0,42,161:39:0 ./.:6:0:32:0,18,56:29:0 ./.:2:0:20:0,6,7:8:0 ./.:2:2:14:49,6,0:27:0 ./.:0:0:0:0, 0,0:.:0 ./.:3:0:23:0,9,59:29:0 0/0:22:0:80:0,66,156:39:0 ./.:3:0:23:0,9,43:26:0

A few lines from the 1,000 geones dataset:

1 10583 rs58108140 G A 100 PASS AVGPOST=0.7707;RSQ=0.4319;LDAF=0.2327;ERATE=0.0161;AN=2184;VT=SNP;AA=.;THETA=0.0046;AC=314;SNPSOURCE=LOWCOV;AF=0.14;ASN_AF=0.13;AMR_AF=0.17;AFR_AF=0.04;EUR_AF=0.21 GT:DS:GL 0|0:0.200:-0.18,-0.47,-2.42 0|0:0.150:-0.24,-0.44,-1.16 0|0:0.150:-0.15,-0.54,-3.12 0|1:0.600:-0.48,-0.48,-0.48

1 13957 rs201747181 TC T 28 PASS AA=TC;AC=35;AF=0.02;AFR_AF=0.02;AMR_AF=0.02;AN=2184;ASN_AF=0.01;AVGPOST=0.8711;ERATE=0.0065;EUR_AF=0.02;LDAF=0.0788;RSQ=0.2501;THETA=0.0100;VT=INDEL GT:DS:GL 0|0:0.050:0,0,0 0|1:0.650:0,0,0 0|0:0.100:0,0,0 0|0:0.350:0,0,0 0|0:0.050:0.00,-0.30,-4.10 0|0:0.650:0,0,0 0|0:0.150:0,0,0 0|0:0.150:0.00,-0.30,-4.10

1 55249 rs200769871 C CTATGG 443 PASS AA=C;AC=151;AF=0.07;AFR_AF=0.03;AMR_AF=0.08;AN=2184;ASN_AF=0.16;AVGPOST=0.9073;ERATE=0.0063;EUR_AF=0.02;LDAF=0.0968;RSQ=0.5891;THETA=0.0038;VT=INDEL GT:DS:GL 0|0:0.000:0.00,-0.30,-6.70 0|0:0.000:0,0,0 0|0:0.200:0,0,0 0|0:0.050:0,0,0 0|1:0.800:0,0,0 0|0:0.000:0.00,-0.30,-6.70

[Zev.Kronenberg]

So after looking at the VCF tools subroutine _format_line_hash I cannot tell you what is wrong. The code spans several pages and is not documented. Grrr - bad coding.

[Adam]

I suggest you ask this question on the vcftools help mailing list, as it is more likely to be seen by the developer of the perl module (vcftools-help (at) lists.sourceforge.net).