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  • bioinfosm
    Senior Member
    • Jan 2008
    • 483

    ChipSEQ on Solexa (low % align, unusable reads)

    Hi,

    I thought there was some discussion, but could not find it - regarding the data output from a chipSEQ run using the Solexa GA.

    There is very little dna to work with, is what the sequencing team says. But after a run through the solexa pipeline, I see under 20% passing-filter reads aligning to the reference genome. That is way too low, and that too with error rate of about 5% !

    Are there any changes or workarounds as to what might be causing it? or its simply blame it on the data! The number of clusters, % pass, intensities all look fine..
    --
    bioinfosm
  • new300
    Member
    • Mar 2008
    • 50

    #2
    How many clusters are you getting per tile and what are the sequences that don't align, are they all the adapter sequence perhaps?

    Comment

    • bioinfosm
      Senior Member
      • Jan 2008
      • 483

      #3
      Originally posted by new300 View Post
      How many clusters are you getting per tile and what are the sequences that don't align, are they all the adapter sequence perhaps?
      Its about 10,000-15,000 PF clusters. I am yet to check match with adapter, is there a dataset I could use to do that? A reference sequence to check for adapter?
      --
      bioinfosm

      Comment

      • bioinfosm
        Senior Member
        • Jan 2008
        • 483

        #4
        And the phiX control looks perfectly fine .. less than 1% error, good data overall..
        --
        bioinfosm

        Comment

        • bioinfosm
          Senior Member
          • Jan 2008
          • 483

          #5
          What should be the way to detect adapter contamination in my chipSEQ data?

          I tried using this as reference set for ELAND, but did not work as expected, any ideas? I would simply want to see what % of reads are adapters or adapter dimers, etc.

          >Adapters1a
          GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG
          >Adapters1b
          ACACTCTTTCCCTACACGACGCTCTTCCGATCT
          --
          bioinfosm

          Comment

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