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Old 06-20-2013, 07:43 AM   #4
Location: Livermore, CA

Join Date: Apr 2011
Posts: 27

I've been doing 16S amplicon sequencing lately and I've found that quantifying by Qubit and pooling works fine, although not quite as good as qPCR. On most runs I see only a slight (+/- 2%) variation in number of reads from any particular barcode indicating that the multiplexing worked well. Given the extra time and cost of running the qPCR, I don't think it's necessary because it only gives slightly better results. For anything other that amplicon sequencing though I would say that quantification by qPCR is an absolute requirement.
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