Hi I'm trying to compute quality metrics for RNA-seq data using RNA-SeQC software, it is asking for read groups of bam file.
I tried using the command samtools view DGL-104.bam | grep '^@RG' but I'm not getting anything. How do I solve this? Shall I add read groups?
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gskbioinfo143 •
Try samtools view -H DGL-104.bam | grep '^@RG' instead. If there are no read groups then yes, you'll need to add them (normally one knows if one has already done this).
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Devon Ryan
Yes I tried this command too there are no read groups. How to add read groups? Any suggestions?
Thanks
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gskbioinfo143 •
Picard Tools has an AddOrReplaceReadGroups command.
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Devon Ryan
Hey I tried adding read groups and did analysis with RNA-SeQC thre was some output but it showed error as mentioned below
Running GATK Depth of Coverage Analysis ....
Arguments: -T DepthOfCoverage -R hg19_.fasta -I DGL-reorder-104.bam -o ./testReport//TestId/highexpr//perBaseDoC.out -L ./testReport//TestId/highexpr/intervals.list -l ERROR
Arguments Array: [-T, DepthOfCoverage, -R, hg19_.fasta, -I, DGL-reorder-104.bam, -o, ./testReport//TestId/highexpr//perBaseDoC.out, -L, ./testReport//TestId/highexpr/intervals.list, -l, ERROR]
Exception in thread "main" java.lang.OutOfMemoryError: GC overhead limit exceeded
at java.util.LinkedList.linkLast(LinkedList.java:140)
at java.util.LinkedList.add(LinkedList.java:336)
at org.broadinstitute.sting.gatk.iterators.LocusIteratorByState$ReadStateManager.addReadsToSample(LocusIteratorByState.java:762)
at org.broadinstitute.sting.gatk.iterators.LocusIteratorByState$ReadStateManager.collectPendingReads(LocusIteratorByState.java:700)
at org.broadinstitute.sting.gatk.iterators.LocusIteratorByState.lazyLoadNextAlignmentContext(LocusIteratorByState.java:375)
at org.broadinstitute.sting.gatk.iterators.LocusIteratorByState.hasNext(LocusIteratorByState.java:345)
at net.sf.picard.util.PeekableIterator.advance(PeekableIterator.java:70)
at net.sf.picard.util.PeekableIterator.next(PeekableIterator.java:57)
at org.broadinstitute.sting.gatk.executive.WindowMaker$WindowMakerIterator.advance(WindowMaker.java:168)
at org.broadinstitute.sting.gatk.executive.WindowMaker$WindowMakerIterator.hasNext(WindowMaker.java:137)
at org.broadinstitute.sting.gatk.datasources.providers.LocusView.advance(LocusView.java:156)
at org.broadinstitute.sting.gatk.datasources.providers.LocusView.hasNextLocus(LocusView.java:122)
at org.broadinstitute.sting.gatk.datasources.providers.AllLocusView.advance(AllLocusView.java:102)
at org.broadinstitute.sting.gatk.datasources.providers.AllLocusView.hasNext(AllLocusView.java:60)
at org.broadinstitute.sting.gatk.traversals.TraverseLoci.traverse(TraverseLoci.java:50)
at org.broadinstitute.sting.gatk.traversals.TraverseLoci.traverse(TraverseLoci.java:18)
at org.broadinstitute.sting.gatk.executive.LinearMicroScheduler.execute(LinearMicroScheduler.java:63)
at org.broadinstitute.sting.gatk.GenomeAnalysisEngine.execute(GenomeAnalysisEngine.java:248)
at org.broadinstitute.sting.gatk.CommandLineExecutable.execute(CommandLineExecutable.java:113)
at org.broadinstitute.sting.commandline.CommandLineProgram.start(CommandLineProgram.java:236)
at org.broadinstitute.sting.commandline.CommandLineProgram.start(CommandLineProgram.java:146)
at org.broadinstitute.cga.rnaseq.gatk.GATKTools.runDoC(GATKTools.java:59)
at org.broadinstitute.cga.rnaseq.PerBaseDoC.runDoC(PerBaseDoC.java:901)
at org.broadinstitute.cga.rnaseq.PerBaseDoC.runDoC(PerBaseDoC.java:870)
at org.broadinstitute.cga.rnaseq.RNASeqMetrics.runMetrics(RNASeqMetrics.java:271)
at org.broadinstitute.cga.rnaseq.RNASeqMetrics.execute(RNASeqMetrics.java:171)
at org.broadinstitute.cga.rnaseq.RNASeqMetrics.main(RNASeqMetrics.java:139)
Please I need suggestion how to solve
Thanks
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gskbioinfo143 •
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