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  • Raw read count to reference genes

    I set a my own reference database containg selected genes (n=1500).
    I had a 454 data about 100MB.

    I run Bowtie 2 to get SAM, sorted/indexed. How do I get a excel-like raw read count output to each mapped reference gene? I mean simple count rather than expression study.

    I noted there are some suggestion in the forum to use Bedtools or HTseq. Are they appropriate for solving my issue. If so, which performs better?

    Thanks a lot.

  • #2
    Originally posted by fanx View Post
    I set a my own reference database containg selected genes (n=1500).
    I had a 454 data about 100MB.

    I run Bowtie 2 to get SAM, sorted/indexed. How do I get a excel-like raw read count output to each mapped reference gene? I mean simple count rather than expression study.

    I noted there are some suggestion in the forum to use Bedtools or HTseq. Are they appropriate for solving my issue. If so, which performs better?

    Thanks a lot.
    Try using BEDTools to intersect your .bam file with a .bed file with gene positions. Then you can process that with cut | uniq, or whatever.

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    • #3
      swbarnes2: Thanks your suggestion. I did find BedTools has such a function. One thing to bother me is how to get coordinating Bed or GFF file. This is because I aligned my reads to NIH viral reference database for which only fasta and gbff are available for the download.

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      • #4
        Take a look at our read counter tool: http://seqanswers.com/forums/showthread.php?p=134850
        https://code.google.com/p/recog/

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