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  • Pooling identical RAD-library PCRs - better for seq of unique fragments?

    Hi, I have two identical RAD-libraries that I PCR'd in two separate reactions (i.e. final PCR before Illumina seq). One is slightly lower concentration, but I wondered if pooling them at some ratio would help with getting unique RAD-tags in my sample. Does anyone have an opinion on this?

    Thanks,
    Rachael
    RAD-Rachael

  • #2
    RAD libraries are generally of high complexity, so there probably isn't any advantage in pooling for reducing sampling bias. There could be a little help from pooling in reducing PCR mutations that become amplified, as the same set of mutations are unlikely to have happened in both pools (but I don't think this is typically a problem anyway).
    Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

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    • #3
      Thanks for your thoughts!
      RAD-Rachael

      Comment

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