picard markdup error:Value was put into PairInfoMap more than once

Hi, all!

I get the error information Value was put into PairInfoMap more than once when I use picard to mark duplication.
I have already tested the newest version bwa 0.7.16a and picard 2.10.7.
My mapping paramater is bwa mem -M.
All paired reads id are unique in fq.

I use samtools view this.bam|grep readsname, the output is captured as this picture:
https://p.qlogo.cn/qqmail_head/LIND77SSexibQw48mEewIK3YKyoKCpB06NW5gSKticJqN2mMnvbd8S7KZdvcuWGj31sIGFzvpM9Bg/0
I have read the pages about this trouble asked before on gatk forum and biostar forum. I think the secondary hits in bam are correct, compared to the previous reported condition, but not for sure. Therefore, I come here to ask for professional help.

Comments

  • SheilaSheila Broad InstituteMember, Broadie, Moderator

    @Dicor
    Hi,

    Sorry for the delay. Have a look at this thread.

    This error usually means a read name occurs twice in your BAM file. Did you by any chance merge two FASTQ files before aligning? Make sure your Read Groups are properly specified. Have a look at this dictionary entry for more help on read groups.

    -Sheila

  • DicorDicor chinaMember

    @Sheila said:
    @Dicor
    Hi,

    Sorry for the delay. Have a look at this thread.

    This error usually means a read name occurs twice in your BAM file. Did you by any chance merge two FASTQ files before aligning? Make sure your Read Groups are properly specified. Have a look at this dictionary entry for more help on read groups.

    -Sheila

    @Sheila
    Thank you for your reply.
    I have already mentioned that all reads ids are unique in my fq files.
    Could you please have a look at the picture?

    I think the problem is both fragments have second hit on ref and are marked. But picard can't recognize them.

  • SkyWarriorSkyWarrior TurkeyMember

    Can you filter your bam file with samtools view -f 0x2 and try marking duplicates again with picard.

    This solved my problem many times with the same error.

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