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  • jmt
    Junior Member
    • Jun 2011
    • 5

    weird output with BWA paired end alignment

    Hi all,

    I'm running into something strange.

    I'm trying to do paired-end alignment with BWA.

    I did the following:

    bwa aln fasta pair1.fastq > pair1.sai
    bwa aln fasta pair2.fastq > pair2.sai

    Then, I did

    bwa sampe fasta pair1.sai pair2.sai pair1.fasta pair2.fastq > bam

    However, when I looked at my bam, I got alignments such as this:

    seq1 117 chr10 49643024 0 * = 49643024 0 CCCGTATAACTCCATTCTTGGTTCTCTCCTCCAAGTCCATAACTTCCTTATATATCAATTCTTTCCACT
    seq1 153 chr10 49643024 37 100M = 49643024 0 AGACAGCAGTCTAGAAGCAGCAGCTGGGCCTCTGGGCTGCTGTAGATGACTACTTGGGCCATCGGGGGG

    My question is: shouldn't seq1 (first in pair; flag 117) have a "*" under column 3, signifying that it's unaligned?

    What is going on here?
  • sudders
    Member
    • Dec 2011
    • 32

    #2
    We've noticed this as well. BWA seems to give a position to unaligned reads.

    Comment

    • jp.
      Senior Member
      • Jul 2013
      • 142

      #3
      How about if give commands like this:
      bwa aln -t 4 -f Sample_R1.sai ./hg19.fasta ./Sample_R1.fastq
      bwa aln -t 4 -f Sample_R2.sai ./hg19.fasta ./Sample_R2.fastq

      Comment

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