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  • quBit fiability for library quant?

    My core facility tells me that libraries should be above 3ng/ul for reliable quantification with the QuBit - mine are at 1ng/ul, which according to the manufacturer is perfectly within the assay range (HS dna assay).

    I was wondering what was the general consensus on low concentration libraries - are they problematic / variable due to poor accuracy in the quantification? Should I quantify them by PCR or just reconcentrate them?

    Thanks

  • #2
    What kind of library are you trying to quantify? We won't run a library that isn't quantified by qPCR prior to loading on the MiSeq. We will quantify certain types of libraries using the Qubit prior to pooling though.

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    • #3
      Its a chip-seq library, so low amount of starting material; adding to that, I'm doing minimal amount of PCR (15 cycles in that case instead of 18) to minimize PCR duplicates, which is important for my specific project. Its to load on a HiSeq - so qPCR is definitively the way to go then?

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      • #4
        We too count on qPCR for final quantification. A qubit is better than a nanodrop but still can be problematic at times.

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