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  • diagen
    Member
    • Jun 2012
    • 13

    #1

    Help! Strange discrepancy in the mapping output

    Hi,

    I analyze the public RNA-seq data GSE29278 from the Ren lab using this pipeline: convert sra to FASTQ - map to mm10 by BWA default - convert sam to bed by SamTools. Remarkably, while the number of FASTQ reads is comparable in all datafilels, about half of them result in 10-20x less mapped reads than in others.
    How to explain this? Could it be a problem of BWA or something else is wrong?
    Will appreciate for any suggestions.

    datafile #FastqReads #mappedReads
    SRR207090.sra 53680296 10916922
    SRR207091.sra 55117644 11007351
    SRR207092.sra 137872756 25021157
    SRR207093.sra 138580112 25982948
    SRR207094.sra 122571612 24707083
    SRR207095.sra 138402388 27051256
    SRR207096.sra 96303468 19404082
    SRR207097.sra 93965080 19379369
    SRR207098.sra 138521432 26517816
    SRR207099.sra 129879756 25531487
    SRR207100.sra 101842656 20724671
    SRR207101.sra 94289428 17374918
    SRR207102.sra 99444472 20386170
    SRR207103.sra 97366804 19698156
    SRR207104.sra 89976804 18662045
    SRR207105.sra 94502252 18551594
    SRR207106.sra 134119212 25478228
    SRR207107.sra 136421136 25550764
    SRR207108.sra 112083680 21283621
    SRR207109.sra 103706068 19259646
    SRR392604.sra 81341016 1167997
    SRR392605.sra 79523572 919123
    SRR392606.sra 107383260 1493933
    SRR392607.sra 108810768 1232292
    SRR392608.sra 80765908 1247112
    SRR392609.sra 78333052 890879
    SRR392610.sra 72429160 1225045
    SRR392611.sra 58510300 1299132
    SRR392613.sra 121652996 1602188
    SRR392614.sra 29990064 205181
    SRR392615.sra 30096232 251824
    SRR392616.sra 96376652 1586269
    SRR392617.sra 79150596 1040652
    SRR392618.sra 22528760 116048
    SRR392619.sra 22024856 124722
  • diagen
    Member
    • Jun 2012
    • 13

    #2
    So the mapping efficiency is around 20% and 1% for these series of datafiles. Actually 20% is not good either.

    Comment

    • diagen
      Member
      • Jun 2012
      • 13

      #3
      The problem was in index sequences that were not removed.
      Solution: remove flanking 8bp or run bowtie in local alignment mode.

      Comment

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