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  • bioinfosm
    Senior Member
    • Jan 2008
    • 483

    #1

    MAQ paired end mapping statistic

    Here is a sample stdout when using maq for paired-end reads.
    # reads mapped in pairs is an odd number. How can that be?
    94% reads mapped, but only 41% mapped in pairs, and about 1% can be rescued by 'moving' the reads. So the remaining 50% of 'mapped' reads, map to different chromosomes?
    Are only the reads mapped in pairs used for SNP calling using downstream MAQ analysis?

    -- 48 potential soa-indels pass the filter.
    -- 3551 potential pe-indels pass the filter.

    -- == statmap report ==

    -- # single end (SE) reads: 0
    -- # mapped SE reads: 0 (/ 0 = NA%)
    -- # paired end (PE) reads: 21532606
    -- # mapped PE reads: 20163428 (/ 21532606 = 93.64%)
    -- # reads that are mapped in pairs: 8326749 (/ 20163428 = 41.29%)
    -- # Q>=30 reads that are moved to meet mate-pair requirement: 899 (/ 8326749 = 0.01%)
    -- # Q<30 reads that are moved to meet mate-pair requirement: 5519 (0.06%)
    --
    bioinfosm
  • MegW
    Junior Member
    • Jun 2009
    • 2

    #2
    bioinfosm, did you ever work out an answer to this? I also have an odd number of reads mapped as pairs and am wondering if it is a sign that something has gone wrong.

    You have probably already worked this other part out, but the answer to your question about whether only pair-mapped reads are used for SNP calling is that it depends on which flags you set. If you use Maq easyrun, I believe that the default setting is to include reads that are not mapped in pairs. You can change this behaviour if you run the scripts yourself, by setting the -p flag for assemble.

    Comment

    • Jonathan
      Member
      • Jun 2009
      • 36

      #3
      Did you have a look at what the 'maq map' step itself told about the mapping?
      (The last view lines of the ... I think stderr-output)
      Perhaps there is some discrepancy when reading the paired-information to produce your output.

      (Btw. how was it generated? I haven't seen this report yet)

      Comment

      • MegW
        Junior Member
        • Jun 2009
        • 2

        #4
        Hi Jonathan,

        Thanks for the suggestion. I've seen this same output consistently for two of my resequencing datasets, but never for a third generated from the same barcoded Illumina lane, no matter whether I map the reads using easyrun or by running the scripts individually (all with default values, except for maq map, which I have run with both default and specificed max insert sizes, and maq assemble, which I have run both with and without -p flag). No matter which way I do it, I always get an odd number of reads mapped in pairs for those two datasets.

        I can't see anything obviously odd in the maq map output, but I am also not totally sure what I am looking for... Here are the tails of two stderr outputs, the first for the well-behaved dataset, and the second for one of the odd-number datasets. Thanks for any insights!


        [ma_load_reads] loading reads...
        [ma_load_reads] 2451785*2 reads loaded.
        [mapping_count_single] 5, 17, 22, 35
        [match_data2mapping] 218 pairs are added afterwards.
        [match_data2mapping] (328642, 316222) reads are moved to meet paired-end requirement.
        [match_data2mapping] quality counts of the first reads: (305, 328337); second reads (265, 315957)
        [cal_insert_size] 624562 read pairs counted. insert size: 262.259648 +/- 26.880905
        [maq_make_aln_candidate] 144376 candidates added
        [maq_indel_pe] 144376 candidates for alignment
        [maq_indel_pe] CPU time: 10.790 sec
        [match_data2mapping] 4447176 out of 4903570 raw reads are mapped with 4217234 in pairs.
        -- (total, isPE, mapped, paired) = (4903570, 1, 4447176, 4217234)




        [ma_load_reads] loading reads...
        [ma_load_reads] 3018849*2 reads loaded.
        [mapping_count_single] 5, 17, 22, 36
        [match_data2mapping] 269 pairs are added afterwards.
        [match_data2mapping] (459993, 455444) reads are moved to meet paired-end requirement.
        [match_data2mapping] quality counts of the first reads: (203, 459790); second reads (168, 455276)
        [cal_insert_size] 811214 read pairs counted. insert size: 247.372625 +/- 17.279953
        [maq_make_aln_candidate] 178357 candidates added
        [maq_indel_pe] 178357 candidates for alignment
        [maq_indel_pe] CPU time: 13.950 sec
        [match_data2mapping] 5856680 out of 6037698 raw reads are mapped with 5625861 in pairs.
        -- (total, isPE, mapped, paired) = (6037698, 1, 5856680, 5625861)

        Comment

        • knott76
          Junior Member
          • Sep 2008
          • 2

          #5
          I'm bumping this back up - hopefully someone has an answer.

          I have a dataset generated from 12 bar-codes in a a 2x50 Illumina lane. Following MAQ Easyrun alignment, I'm getting a similar error, and wondering if anyone has a solution. Any suggestions would be greatly appreciated.

          The stat report for one of the bar-coded samples looks like the following:

          -- == statmap report ==

          -- # single end (SE) reads: 0
          -- # mapped SE reads: 0 (/ 0 = NA%)
          -- # paired end (PE) reads: 1985630
          -- # mapped PE reads: 1805746 (/ 1985630 = 90.94%)
          -- # reads that are mapped in pairs: 92591 (/ 1805746 = 5.12%)
          -- # Q>=30 reads that are moved to meet mate-pair requirement: 5 (/ 92591 = 0%)
          -- # Q<30 reads that are moved to meet mate-pair requirement: 14 (0.01%)

          Comment

          • Kasycas
            Member
            • Sep 2009
            • 22

            #6
            I have the same problem

            -- == statmap report ==

            -- # single end (SE) reads: 0
            -- # mapped SE reads: 0 (/ 0 = NA%)
            -- # paired end (PE) reads: 30130376
            -- # mapped PE reads: 28705774 (/ 30130376 = 95.27%)
            -- # reads that are mapped in pairs: 7988607 (/ 28705774 = 27.82%)
            -- # Q>=30 reads that are moved to meet mate-pair requirement: 660 (/ 7988607 = 0%)
            -- # Q<30 reads that are moved to meet mate-pair requirement: 76318 (0.95%)


            Very confusing!

            Anyone solve this issue?

            Thanks.

            Comment

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