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  • xiang
    Member
    • Mar 2009
    • 13

    #1

    maq problem for 108bp reads.

    I just started to use maq to map the paired-end reads of 108bp.
    When I use command "maq map -e 200 -a 500 a1.bfq a2.bfq",
    I always got this:

    -- maq-0.7.1
    [ma_load_reads] loading reads...
    [ma_load_reads] set length of the first read as 108.
    [ma_load_reads] set length of the second read as 108.
    [ma_load_reads] 1456472*2 reads loaded.
    [ma_longread2read] encoding reads... 2912944 sequences processed.
    [ma_match] set the minimum insert size as 109.
    [match_core] Total length of the reference: 1586004629
    [match_core] round 1/3...
    [match_core] making index...
    [match_index_sorted] no reasonable reads are available. Exit!

    I checked the data using "maq bfq2fastq a1.bfq". It shows the data is as this:

    @IL41_3804:3:10:236:221/1
    CAGTGATTTCGTCATTTTTCAAGTCGTTAAGTGGATATTCCTCATTTTCCATGATTTTCAGTTTTCTTGCCATATTCCATGTCCCACAGTGGACATTTCTAAATTTTC
    +
    CCDCDBACBABE<CCDDDCCACA;=DCCCBCACCBCCCA?BB=BCCCB@CACA<CCCC?@B@@CC?CB@<=@@0-3<6=A@@>77;5978A?<;=@?@=A?;:=8;??

    I can't find any problem. Anyone has idea about this? Thanks.
  • suseq
    Junior Member
    • Sep 2009
    • 3

    #2
    I got the same problem with 75 bp long paired end reads. Do you find a resolution?

    Comment

    • nilshomer
      Nils Homer
      • Nov 2008
      • 1283

      #3
      Originally posted by xiang View Post
      I just started to use maq to map the paired-end reads of 108bp.
      When I use command "maq map -e 200 -a 500 a1.bfq a2.bfq",
      I always got this:

      -- maq-0.7.1
      [ma_load_reads] loading reads...
      [ma_load_reads] set length of the first read as 108.
      [ma_load_reads] set length of the second read as 108.
      [ma_load_reads] 1456472*2 reads loaded.
      [ma_longread2read] encoding reads... 2912944 sequences processed.
      [ma_match] set the minimum insert size as 109.
      [match_core] Total length of the reference: 1586004629
      [match_core] round 1/3...
      [match_core] making index...
      [match_index_sorted] no reasonable reads are available. Exit!

      I checked the data using "maq bfq2fastq a1.bfq". It shows the data is as this:

      @IL41_3804:3:10:236:221/1
      CAGTGATTTCGTCATTTTTCAAGTCGTTAAGTGGATATTCCTCATTTTCCATGATTTTCAGTTTTCTTGCCATATTCCATGTCCCACAGTGGACATTTCTAAATTTTC
      +
      CCDCDBACBABE<CCDDDCCACA;=DCCCBCACCBCCCA?BB=BCCCB@CACA<CCCC?@B@@CC?CB@<=@@0-3<6=A@@>77;5978A?<;=@?@=A?;:=8;??

      I can't find any problem. Anyone has idea about this? Thanks.
      There is a maximum read length. You'll have to dig in the source code. Try BWA, the successor to MAQ.

      Comment

      • Jonathan
        Member
        • Jun 2009
        • 36

        #4
        Originally posted by suseq View Post
        I got the same problem with 75 bp long paired end reads. Do you find a resolution?
        Ah, but MAQ works for 76bp PE - at least the latest version did for my dataset.
        best
        -Jonathan

        Comment

        • xiang
          Member
          • Mar 2009
          • 13

          #5
          I have tried more data. Now I am sure that maq can work for 108bp reads since I have already got some successful cases.

          But for some data, It just reports this error. I don't know why.

          Comment

          • suseq
            Junior Member
            • Sep 2009
            • 3

            #6
            The newest version is MAQ 0.7.1, or not? I tried to use this version but get always:
            "[match_index_sorted] no reasonable reads are available. Exit! "
            The rest looks similar to Xiang`s example: the reads were loaded, the reference as well, but than it stopps.

            Do you have any idea why???

            Comment

            • stephlg
              Junior Member
              • Nov 2008
              • 2

              #7
              some news?

              In my lab, we sequenced 4 samples on a PE 2x60.All samples have been sequenced on the same flowcell and sequences have been generated with the same pipeline.

              I used maq and I managed to align only 2 out of 4 samples. For the two non aligned samples I got the same error as you had.
              I tried different parameters but I always have the same error.

              I really don't understand why I couldn't align 2 samples since all samples have been processed the same way! They are in the same iFastq format.

              Some news about this issue?
              Last edited by stephlg; 02-01-2010, 08:32 AM.

              Comment

              • lh3
                Senior Member
                • Feb 2008
                • 686

                #8
                This is most likely that your reads have too many 'N's.

                Comment

                • xiang
                  Member
                  • Mar 2009
                  • 13

                  #9
                  Basically, this part of data is of low quality.

                  What I have done for them is: chopping them down to a smaller read-length.
                  for example 108bp -> 93bp and then align it. You can use maq -1 93 to do this without changing the .bfq files.

                  Comment

                  • stephlg
                    Junior Member
                    • Nov 2008
                    • 2

                    #10
                    Actually I had a look on my data and it appeared that a lot of sequences contained a lot of Ns. So I filtered out tags with Ns and aligned them. Finally it has worked.

                    Thank you for your help

                    Comment

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