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  • jdpr_100
    Member
    • Jun 2014
    • 14

    #1

    Why BWA output didn't agree with bowtie2

    I am working on the alignment between single-end read and reference genome using BWA and bowtie2. I got different results from BWA and bowtie. I couldn’t figure out what I am doing wrong.
    bowtie2 commands:
    bowtie2 --sensitive-local -x <indexed ref> -q <reads> -S output.sam
    I got following statistics with bowtie2.
    total reads 411526452
    69195599 (16.81%) aligned 0 times
    304002593 (73.87%) aligned exactly 1 time
    38328260 (9.31%) aligned >1 times
    I also check the XM tag in .sam file generated from bowtie2
    31720802 XM:i:5
    46327246 XM:i:4
    58292573 XM:i:3
    59969734 XM:i:2
    46552300 XM:i:1
    22355229 XM:i:0
    When I run bwa with the following commands, I would like to generate similar statistics.
    bwa index ref.fa
    bwa aln –n 5 ref.fa reads.fq > out.sai
    bwa samse ref.fa out.sai reads.fq > out.sam
    And , get some stats with following commands.
    samtools view -c output.sorted.bam
    samtools view -c -q1 output.sorted.bam
    samtools view -c -F 4 output.bam
    samtools view -c -f 4 output.bam
    total reads 411526452
    # of reads uniquely mapped 179363510 43.58%
    unmapped reads 230229393 55.94%
    mapped reads 181297059 44.05%
    I also check the XM tag in output.sam generated from bwa.
    18855884 XM:i:5
    32102066 XM:i:4
    41821781 XM:i:3
    43223159 XM:i:2
    31594809 XM:i:1
    13301580 XM:i:0
    There are huge different between the results from BWA and that from Bowtie2. Only about 44% reads have been aligned with the ref using bwa.
    What is the reason cause this difference?
    Thank your for your help in advance
    Last edited by jdpr_100; 06-25-2014, 07:26 AM.
  • dpryan
    Devon Ryan
    • Jul 2011
    • 3478

    #2
    Why would you expect them to be similar? Different algorithms will always produce different results.

    Comment

    • jdpr_100
      Member
      • Jun 2014
      • 14

      #3
      Compared to 71% in Bowtie2, only 44% reads can be mapped to reference genome using BWA. I expect at least 60%. I am not sure whether the results are normal or not. Or, there something wrong with -n 5 (mismatches) in BWA.

      Comment

      • Brian Bushnell
        Super Moderator
        • Jan 2014
        • 2709

        #4
        Clearly, the problem here is that you're not using BBMap

        But when using bwa, you should run bwa-mem instead of bwa aln, which is slower and cannot handle as many errors. Bwa-mem will give you output that is more similar to bowtie2.

        Comment

        • jdpr_100
          Member
          • Jun 2014
          • 14

          #5
          Hi Brian Bushnell, Thank you for your suggestion. In my case, reads length range from 50 to 100bp. Can bwa-mem handle 50-100bp reads?

          Comment

          • Brian Bushnell
            Super Moderator
            • Jan 2014
            • 2709

            #6
            Yes, it can.

            Comment

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