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  • 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

  • #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


    • #3
      change Enzyme

      Comment


      • #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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