Tagged with #variants
1 documentation article | 0 events or announcements | 3 forum discussions


1. What file formats do you support for variant callsets?

We support the Variant Call Format (VCF) for variant callsets. No other file formats are supported.

2. How can I know if my VCF file is valid?

VCFTools contains a validation tool that will allow you to verify it.

3. Are you planning to include any converters from different formats or allow different input formats than VCF?

No, we like VCF and we think it's important to have a good standard format. Multiplying formats just makes life hard for everyone, both developers and analysts.

Sorry, there are no publicly available documents of this type with the tag #variants. Try one of the other types.

Hi,

Could you tell me when we can use new version of SnpEff with GATK?

I have some bugs :

ERROR ------------------------------------------------------------------------------------------
ERROR A USER ERROR has occurred (version 2.0-35-g2d70733):
ERROR The invalid arguments or inputs must be corrected before the GATK can proceed
ERROR Please do not post this error to the GATK forum
ERROR
ERROR See the documentation (rerun with -h) for this tool to view allowable command-line arguments.
ERROR Visit our website and forum for extensive documentation and answers to
ERROR commonly asked questions http://www.broadinstitute.org/gatk
ERROR
ERROR MESSAGE: Could not create module ActiveRegionBasedAnnotation because BUG: Cannot find expected constructor for class caused by exception org.broadinstitute.sting.gatk.walkers

.annotator.interfaces.ActiveRegionBasedAnnotation.()

ERROR ------------------------------------------------------------------------------------------
ERROR ------------------------------------------------------------------------------------------
ERROR A USER ERROR has occurred (version 2.0-35-g2d70733):
ERROR The invalid arguments or inputs must be corrected before the GATK can proceed
ERROR Please do not post this error to the GATK forum
ERROR
ERROR See the documentation (rerun with -h) for this tool to view allowable command-line arguments.
ERROR Visit our website and forum for extensive documentation and answers to
ERROR commonly asked questions http://www.broadinstitute.org/gatk
ERROR
ERROR MESSAGE: Could not create module ExperimentalAnnotation because BUG: Cannot find expected constructor for class

caused by exception org.broadinstitute.sting.gatk.walkers.annotator.interfaces.ExperimentalAnnotation.()

ERROR ------------------------------------------------------------------------------------------
ERROR ------------------------------------------------------------------------------------------
ERROR A USER ERROR has occurred (version 2.0-35-g2d70733):
ERROR The invalid arguments or inputs must be corrected before the GATK can proceed
ERROR Please do not post this error to the GATK forum
ERROR
ERROR See the documentation (rerun with -h) for this tool to view allowable command-line arguments.
ERROR Visit our website and forum for extensive documentation and answers to
ERROR commonly asked questions http://www.broadinstitute.org/gatk
ERROR
ERROR MESSAGE: Could not create module RankSumTest because BUG: cannot instantiate class: must be concrete class caused by exception null
ERROR ------------------------------------------------------------------------------------------

I don't know if I forget some other options linked these annotations. These options are important for me. So I deleted them but if somebody want to use them ...

Regards,

Tiphaine

HI

I am using the following set of commands on GATK2.1.13 to generate a VCF file

echo `java -Xmx20g -jar /usr/bin/GenomeAnalysisTK.jar -I B2_with_ReadGroup.ddup.sorted.bam -R human_g1k_v37.fasta -T RealignerTargetCreator  -o my.intervals -et NO_ET -K /root/sandbox/saket.kumar_iitb.ac.in.key`
echo "Realignment Done at `date`"
echo "Starting IndelRealigner at `date`"

echo `java -Xmx20g -jar /usr/bin/GenomeAnalysisTK.jar -I B2_with_ReadGroup.ddup.sorted.bam -R human_g1k_v37.fasta -T IndelRealigner -targetIntervals my.intervals -o myrealignedBam.bam  -et NO_ET -K /root/sandbox/saket.kumar_iitb.ac.in.key`
echo "Realignment done at `date`"
echo "Starting UnifiedGenotyper at `date`"
echo `java -Xmx20g -jar /usr/bin/GenomeAnalysisTK.jar -l INFO -R human_g1k_v37.fasta -T UnifiedGenotyper    -I myrealignedBam.bam    -o mygatk_vcf.vcf    --output_mode EMIT_ALL_SITES -et NO_ET -K /root/sandbox/saket.kumar_iitb.ac.in.key`
echo "Gentoypxing complete at `date`"

When i do a 'mpileup' for B2_with_ReadGroup.ddup.sorted.bam , I get a devcent 10 MB VCF file. But on the last ste of the above pipeline, my " mygatk_vcf.vcf " is goinging into 81GBs !!

Do you know what is wrong ?

Dear, GATK team, I have done raw snp and indel calling with UnifiedGenotyper following the command line below.

java -Xmx16g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -glm BOTH -R ucsc.hg19.fasta -T UnifiedGenotyper -I ERR031029.marked.realigned.fixed.recal.bam -I ERR031030.marked.realigned.fixed.recal.bam -D dbsnp_135.hg19.vcf -o ERR031030.raw.snps.indels.vcf -metrics snps.metrics -stand_call_conf 50.0 -stand_emit_conf 10.0 -dcov 1000

After that, I did snp filteration using the following command lines.

java -Xmx8g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -R ucsc.hg19.fasta -T SelectVariants --variant ERR031030.raw.snps.indels.vcf -o ERR031030.snpsonly.vcf -selectType SNP

java -Xmx8g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -R ucsc.hg19.fasta -T SelectVariants --variant ERR031030.raw.snps.indels.vcf -o ERR031030.indelsonly.vcf -selectType INDEL

java -Xmx8g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -T VariantRecalibrator -R ucsc.hg19.fasta -input ERR031030.snpsonly.vcf -resource:hapmap,known=false,training=true,truth=true,prior=15.0 hapmap_3.3.hg19.vcf -resource:omni,known=false,training=true,truth=false,prior=12.0 1000G_phase1.indels.hg19.vcf -resource:dbsnp,known=true,training=false,truth=false,prior=6.0 dbsnp_135.hg19.vcf -an QD -an HaplotypeScore -an MQRankSum -an ReadPosRankSum -an MQ -mode SNP -recalFile ERR031030.snp.recal.vcf -tranchesFile ERR031030.snp.tranches.vcf -rscriptFile ERR031030.plots.R

java -Xmx8g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -R ucsc.hg19.fasta -T ApplyRecalibration -input ERR031030.snpsonly.vcf -tranchesFile ERR031030.snp.tranches.vcf -recalFile ERR031030.snp.recal.vcf -o ERR031030.snps.filtered.vcf

java -Xmx16g -jar GenomeAnalysisTK-2.3-9-ge5ebf34/GenomeAnalysisTK.jar -R ucsc.hg19.fasta -T VariantFiltration --variant ERR031030.snps.filtered.vcf -o ERR031030.final.filtered.vcf --filterName "Nov28filters && QD < 2.0 && ReadPosRankSum < -8.0 && MQ < 40.0 && FS > 60.0 && MQRandkSum < -12.5"  --filterExpression "HaplotypeScore > 13.0"

The filtered snp.vcf file came up, however, it seems it contains some problem.

chrM    311     .       T       C       429.19  Nov28filters **_&& QD < 2.0 && ReadPosRankSum < -8.0 && MQ < 40.0 && FS > 60.0 && MQRandkSum < -12.5;VQSRTrancheSNP99.90to100.00   AC=1;AF=0.250;AN=4;BaseQRankSum=-13.010;DP=2000;Dels=0.00;FS=50.500;HaplotypeScore=382.2016;MLEAC=1;MLEAF=0.250;MQ=50.86;MQ0=0;MQRankSum=1.458;QD=0.43;ReadPosRankSum=-10.687;VQSLOD=-6.143e+02;culprit=HaplotypeScore  GT:AD:DP:GQ:PL  0/0:634,353:949:99:0,232,7697   0/1:463,521:945:99:459,0,4190
chrM    410     .       A       T       64750.20        PASS    AC=4;AF=1.00;AN=4;DP=2000;Dels=0.00;FS=0.000;HaplotypeScore=7.3762;MLEAC=4;MLEAF=1.00;MQ=56.04;MQ0=0;QD=32.38;VQSLOD=2.27;culprit=HaplotypeScore        GT:AD:DP:GQ:PL  1/1:0,998:998:99:32010,2926,0   1/1:0,999:999:99:32767,2912,0
chrM    711     .       G       A       62989.20        PASS    AC=4;AF=1.00;AN=4;BaseQRankSum=2.500;DP=2000;Dels=0.00;FS=3.751;HaplotypeScore=8.7084;MLEAC=4;MLEAF=1.00;MQ=56.74;MQ0=1;MQRankSum=-0.107;QD=31.49;ReadPosRankSum=-2.169;VQSLOD=2.46;culprit=HaplotypeScore
      GT:AD:DP:GQ:PL  1/1:0,998:972:99:30899,2808,0   1/1:3,997:972:99:32117,2830,0
chrM    1121    .       T       C       16719.20        Nov28filters && QD < 2.0 && ReadPosRankSum < -8.0 && MQ < 40.0 && FS > 60.0 && MQRandkSum < -12.5;VQSRTrancheSNP99.90to100.00   AC=4;AF=1.00;AN=4;BaseQRankSum=-0.239;DP=2000;Dels=0.00;FS=2.141;HaplotypeScore=22.9003;MLEAC=4;MLEAF=1.00;MQ=21.32;MQ0=703;MQRankSum=-1.627;QD=8.36;ReadPosRankSum=-0.027;VQSLOD=-4.195e+00;culprit=HaplotypeScore     GT:AD:DP:GQ:PL  1/1:3,985:986:99:9547,976,0     1/1:4,983:983:99:7199,739,0
chrM    2489    .       A       C       34.19   LowQual;Nov28filters && QD < 2.0 && ReadPosRankSum < -8.0 && MQ < 40.0 && FS > 60.0 && MQRandkSum < -12.5       AC=1;AF=0.250;AN=4;BaseQRankSum=-17.321;DP=2000;Dels=0.00;FS=180.208;HaplotypeScore=18.7245;MLEAC=1;MLEAF=0.250;MQ=46.52;MQ0=31;MQRankSum=3.365;QD=0.03;ReadPosRankSum=-4.198   GT:AD:DP:GQ:PL  0/1:278,719:950:64:64,0,4623    0/0:309,688:950:99:0,263,6065

For the filter option, most of the filtered snps show Nov28filters rather than PASS or LowQual, what's wrong with that, Are there some problems with my command lines? Thank you so much for your reply.