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  • BWA output bitwise flag for mapped/unmapped reads

    Hi everybody,

    I have a question on a strange alignment I got with BWA (0.5.9). I have come up with a minimal test sequence



    @5:84:16665:11131:Y
    AGAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAAGAGG
    +
    fdffffff]cffff[aa_]aPVVSYQaa\afc`cffcfffdWf`fadaWd`]aZ^TV]X_TaSWaQ[Z



    which apparently can map to the drosophila genome (dm3/R5).

    and has the following SAM output




    5:84:16665:11131:Y 4 chr2LHet 368806 11 68M * 0 0 AGAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAACAGAGAAGAGG GEGGGGGG>DGGGG<BB@>B1774:2BB=BGDADGGDGGGE8GAGBEB8EA>B;?57>9@5B48B2<; XT:A:U NM:i:1 X0:i:1 X1:i:15 XM:i:1 XO:i:0 XG:i:0 MD:Z:1C66




    what baffles me is the bitwise flag = 4, which indicates that this read is unmapped.

    Anybody has any idea what is happening here?

    Thank you,

    Wen

  • #2
    I think I have found the answer in this post

    Discussion of next-gen sequencing related bioinformatics: resources, algorithms, open source efforts, etc


    It turns out the alignment was at the end of the chromosome and contained a overhang which apparently mapped to the next chromosome.

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