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  • mRNA seq 75bp splice file

    Illumina provides a human.49.splice ELAND_RNA_GENOME_SPLICE file for 50bp reads. Does someone have the same file for 75bp reads?

    There is this thought that 75bp reads are too long for mrna-seq, as they may cover more than one splice sites, thus causing more harm than help. Do people have comments on that? We have 75 bp reads and wish to use the Illumina Genome Studio pipeline but do not have access to the 75bp splice file.

    ..
    --
    bioinfosm

  • #2
    use TopHat, it breaks the 75 bp read into 3 segments. I heard TopHat does this to prevent the problem you mentionned, but I am not sure!
    I hope Cole comments on this!

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    • #3
      I'm also wondering about how TopHat handles these smaller segments. If the segment spans across a splice junction, can Bowtie align this or is this information lost?

      Update: just re-read the paper and it explains how it works quite nicely
      Last edited by kch; 09-30-2009, 08:13 PM. Reason: answered my own question

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      • #4
        One of the perl scripts that comes with the pipeline software can be used to generate splice junction indexes of any size you'd like.

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        • #5
          Originally posted by graveley View Post
          One of the perl scripts that comes with the pipeline software can be used to generate splice junction indexes of any size you'd like.
          Could you give the name of that script?
          thanks
          --
          bioinfosm

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