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Old 05-22-2018, 10:19 AM   #34
idedios
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Location: Irvine, CA

Join Date: Mar 2014
Posts: 18
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Quote:
Originally Posted by korostin View Post
1. are you sure, OneTaq Mix is tolerant to U?
2. I'd add size-select step after 1st PCR to solve primer dimers problem (especially if starting with small amounts of DNA)
I checked and OneTaq is compatible with uracil templates. They use it for their bisulfite protocol.

Too bad OneTaq is not a HiFi enzyme. For any liquid biopsy assay where I would want to make calls around 0.1% I would definitely need a HiFi enzyme. Too bad I couldn't find one that is compatible with uracil templates. On the upside OneTaq claims to have 2x higher fidelity than regular Taq.

Size selection after 1st PCR should not be necessary with the FuPa digestion which should digest all primer sequences including template incorporated ones and dimers.

Last edited by idedios; 05-22-2018 at 10:24 AM.
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