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  • chen
    Member
    • Sep 2009
    • 14

    Mate pair, high GC

    It shows that high GC samples may have a lower concentration after PCR.
    Is there any notes to take when preparing the mate pair library with the HIGH GC samples, except the PCR process(increase the denature time).
    Any suggestion would be help. Thanks.
    Cheers,
    Tanya
  • nextgen
    Member
    • Feb 2010
    • 26

    #2
    Can you try betaine, DMSO to lower Tm?

    Originally posted by chen View Post
    It shows that high GC samples may have a lower concentration after PCR.
    Is there any notes to take when preparing the mate pair library with the HIGH GC samples, except the PCR process(increase the denature time).
    Any suggestion would be help. Thanks.

    Comment

    • grace033
      Junior Member
      • May 2010
      • 5

      #3
      change Enzyme

      Comment

      • Michael.James.Clark
        Senior Member
        • Apr 2009
        • 207

        #4
        I guess you're using Phusion HF for your library prep PCR normally. There is a high-GC version of Phusion you could try instead.
        Mendelian Disorder: A blogshare of random useful information for general public consumption. [Blog]
        Breakway: A Program to Identify Structural Variations in Genomic Data [Website] [Forum Post]
        Projects: U87MG whole genome sequence [Website] [Paper]

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