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  • PCR based test for TruSeq library contamination in Pre-PCR lab?

    Hi,
    I am looking for a simple PCR based assay to evaluate if there is TruSeq amplified library present in our Pre-PCR lab. I would like to use the Bioanalyzer to quantitate. Would something as simple as designing primers to the P5/P7 and amplifying work? Could I just use the leftover primers from the library prep kit? How many cycles would you recommend? Any input or ideas welcomed.

  • #2
    Your pre-PCR libraries probably contain doubly ligated product which in itself can amplify. How about running qPCR with some PCR amplified positive controls and comparing Ct values. I can't think of a good way for you to do with with a bioanalyzer. Are you having contamination issues?

    - Genohub

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    • #3
      I use Kapa's qPCR quantitation kit (which as you said is P5/P7 primers). Bioanalyzer won't be sensitive enough.

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