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  • Seqwork
    Junior Member
    • Mar 2014
    • 4

    Low library yield

    Hello Everyone,
    I am new person to RNA Seq work.I made libraries from FFPE RNA as per protocol.RNA input was from 0.5-1ug for different samples.
    But i can hardly see any yield on bioanalyzer and nanodrop readings are also 10ng/ul or below.
    This is disappointing for me.I have been careful.I don't know where i lost my nucleic acid.One thing which i can remember about Ribosomal RNA removal beads step ,those beads were really difficult to dissolve.
    Any suggestions will be welcomed.
    Thanks
  • kwaraska
    Senior Member
    • Nov 2008
    • 131

    #2
    10ng/ul for an RNAseq library is not low-it is in fact quite good.

    What does your bioanalyzer trace show?

    Comment

    • Seqwork
      Junior Member
      • Mar 2014
      • 4

      #3
      I made libraries twice.Both time nanodrop reading were 10ng/ul or below.
      But i checked my libraries only first time on bioanalyzer, and i can see something at right size around 230-260 but very small peaks.
      I have not checked my libraries second time at bioanalyzer, as i thought again i will not see anything.
      someone suggested me that i should see at least 30ng/ul at nanodrop, but obviously bioanalyzer is more accurate.
      what can i do to improve my yield.As i m planning to start it again.
      Thank you for replying.

      Comment

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