PCR woes

schupp at nauvax.ucc.nau.edu schupp at nauvax.ucc.nau.edu
Fri Aug 5 07:25:36 EST 1994

A couple of suggestions:

        BRL suggests that lowering the reaction volume and increasing
        dNTP concentration (400uM max) improves large fragment amp.

        How are you storing your primers?  We've had problems with storing
        10X stocks, moreso when they experience multiple freeze thaw cycles.
        I suggest storing higher concentrations at 4C, making up 10X
        stocks fresh before each PCR run.

        You might try reoptimizing again (primer and Mg++ concentration

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