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  • FerdinandvM
    Junior Member
    • Jul 2013
    • 2
    #1

    ScriptSeq - 2nd peak @ 550bp

    Hi
    I have been preparing libraries from approx 400ng total RNA, mRNA enrichment, using the ScriptSeqV2 kit.
    At the end I amplify with 12 cycles and I get a profile with a 2nd peak around 550bp. I have not been able to get an explanation for this yet.

    If somebody has an idea I would be grateful.

    Thanks Ferdinand

    P.S. Profile attached.
    Attached Files
    Tags: problem, scriptseq
    • Olaf Blue
      Member
      • Nov 2010
      • 58
      #2
      Did you quantify the input RNA for the Scriptseq library? The kit works best with 500 pg to 50 ng input.

        Comment

        • FerdinandvM
          Junior Member
          • Jul 2013
          • 2
          #3
          No, but assuming that mRNA will never be more than 10% of the total RNA I should be in range.
          Could it be an issue with the fragmentation?

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