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  • #1

    Going from biotinylated primer to unlabeled primer in emPCR

    Hello,

    Merely making an observation and having a question. Does anyone know why 454 went from including a biotinylated primer A in their GS20 workflow to current Titanium chemistry (may have been in FLX but I'm not certain) that uses an unlabeled primer in emPCR?

    Current procedure seems quite cumbersome. After emPCR, one has to break emulsion, melt off complementary strand, annealing enrich primer, enrich, melt off enrich primer A, annealing seq primer followed by pyrosequencing. This is all because they no longer use a biotinylated primer in emPCR.

    Any feedback is appreciated.
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