4

I doing the alignment of samples following the GATK pipeline, and doing the MergeBamAlignment,like this:

MergeBamAlignment \
  -ALIGNED $path/file.unsorted.bam \
  -UNMAPPED $path/file.unmapped.bam \
  -O $path/file.merged.bam \
  -R $references/GRCh38.primary_assembly.genome.fa \
  -SO coordinate \
  -TMP_DIR $path/tmp/

I got this error:

Exception in thread "main" java.lang.IllegalArgumentException: Do not use this function to merge dictionaries with different sequences in them. Sequences must be in the same order as well. Found [] and [chr1, chr2, chr3, chr4, chr5, chr6, chr7, chr8, chr9, chr10, chr11, chr12, chr13, chr14, chr15, chr16, chr17, chr18, chr19, chr20, chr21, chr22, chrX, chrY, chrM, GL000008.2, GL000009.2, GL000194.1, GL000195.1, GL000205.2, GL000208.1, GL000213.1, GL000214.1, GL000216.2, GL000218.1, GL000219.1, GL000220.1, GL000221.1, GL000224.1, GL000225.1, GL000226.1, KI270302.1, KI270303.1, KI270304.1, KI270305.1, KI270310.1, KI270311.1, KI270312.1, KI270315.1, KI270316.1, KI270317.1, KI270320.1, KI270322.1, KI270329.1, KI270330.1, KI270333.1, KI270334.1, KI270335.1, KI270336.1, KI270337.1, KI270338.1, KI270340.1, KI270362.1, KI270363.1, KI270364.1, KI270366.1, KI270371.1, KI270372.1, KI270373.1, KI270374.1, KI270375.1, KI270376.1, KI270378.1, KI270379.1, KI270381.1, KI270382.1, KI270383.1, KI270384.1, KI270385.1, KI270386.1, KI270387.1, KI270388.1, KI270389.1, KI270390.1, KI270391.1, KI270392.1, KI270393.1, KI270394.1, KI270395.1, KI270396.1, KI270411.1, KI270412.1, KI270414.1, KI270417.1, KI270418.1, KI270419.1, KI270420.1, KI270422.1, KI270423.1, KI270424.1, KI270425.1, KI270429.1, KI270435.1, KI270438.1, KI270442.1, KI270448.1, KI270465.1, KI270466.1, KI270467.1, KI270468.1, KI270507.1, KI270508.1, KI270509.1, KI270510.1, KI270511.1, KI270512.1, KI270515.1, KI270516.1, KI270517.1, KI270518.1, KI270519.1, KI270521.1, KI270522.1, KI270528.1, KI270529.1, KI270530.1, KI270538.1, KI270539.1, KI270544.1, KI270548.1, KI270579.1, KI270580.1, KI270581.1, KI270582.1, KI270583.1, KI270584.1, KI270587.1, KI270588.1, KI270589.1, KI270590.1, KI270591.1, KI270593.1, KI270706.1, KI270707.1, KI270708.1, KI270709.1, KI270710.1, KI270711.1, KI270712.1, KI270713.1, KI270714.1, KI270715.1, KI270716.1, KI270717.1, KI270718.1, KI270719.1, KI270720.1, KI270721.1, KI270722.1, KI270723.1, KI270724.1, KI270725.1, KI270726.1, KI270727.1, KI270728.1, KI270729.1, KI270730.1, KI270731.1, KI270732.1, KI270733.1, KI270734.1, KI270735.1, KI270736.1, KI270737.1, KI270738.1, KI270739.1, KI270740.1, KI270741.1, KI270742.1, KI270743.1, KI270744.1, KI270745.1, KI270746.1, KI270747.1, KI270748.1, KI270749.1, KI270750.1, KI270751.1, KI270752.1, KI270753.1, KI270754.1, KI270755.1, KI270756.1, KI270757.1].
    at htsjdk.samtools.SAMSequenceDictionary.mergeDictionaries(SAMSequenceDictionary.java:340)
    at picard.sam.SamAlignmentMerger.getDictionaryForMergedBam(SamAlignmentMerger.java:197)
    at picard.sam.AbstractAlignmentMerger.mergeAlignment(AbstractAlignmentMerger.java:384)
    at picard.sam.SamAlignmentMerger.mergeAlignment(SamAlignmentMerger.java:181)
    at picard.sam.MergeBamAlignment.doWork(MergeBamAlignment.java:368)
    at picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:309)
    at picard.cmdline.PicardCommandLine.instanceMain(PicardCommandLine.java:103)
    at picard.cmdline.PicardCommandLine.main(PicardCommandLine.java:113)

srun: error: compute-12: task 0: Exited with exit code 1

Before I also confirmed that the files that I'm trying to merge, are with the same reference sequences. So I don't understand what could be the problem.

terdon
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Rita Soares
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  • Please clarify your specific problem or provide additional details to highlight exactly what you need. As it's currently written, it's hard to tell exactly what you're asking. – Community Aug 21 '23 at 00:45
  • Welcome @RitaSoares. Firstly is this a human genome? Secondly, what have you done unto this point, ie. how did you create the different alignments, especially what were being used? – M__ Aug 21 '23 at 00:49
  • Not sure if this is related, but do you get the same error after sorting your aligned bam file? Also, just checking that you aligned the reads to the same reference ($references/GRCh38.primary_assembly.genome.fa) – jared_mamrot Aug 23 '23 at 06:14
  • @jared_mamrot yes but i got a different error after sorting the aligned bam files, with SortSamSpark: "org.broadinstitute.hellbender.tools.spark.pipelines. SortSamSpark done. Elapsed time: 0.07 minutes.Runtime.totalMemory()=2854748160 htsjdk.samtools.util.RuntimeEOFException: Premature EOF. Expected 4 but only received 0; BinaryCodec in readmode; streamed file (filename not available)". But according to this error that happend to me before, normally i put in the code of MergeBamAlignment the argument "SO coordinate"and this corrects my problem but with this samples doesn't happen that. – Rita Soares Aug 23 '23 at 14:29
  • Sounds to me like your sample might be corrupted/truncated. What does picard ValidateSamFile output for that sample? – jared_mamrot Aug 23 '23 at 23:07
  • I also think that is the case... i will try to do ValidateSamFile to make sure. Thank for your answer – Rita Soares Aug 28 '23 at 14:44

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