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Problems with Bisulfite PCR - (Jul/08/2008 )

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I tried a set of primers with 3 metCs, but they did not work very well. I have designed 5 primer sets so far and these are the only ones which seem to work. I ran a 2 cycle PCR along with it and it seems to have quite a bit of no-specific priming. Is this normal? Anyway to get rid of it????

-DazedNConfused-

Also.... My reverse primer is also methylated as the next base at the 3' is G.

-DazedNConfused-

you would need to optimise your annealing temperature in your PCR I would say


-methylnick-

Thanks! I think I finally got it working!

-DazedNConfused-

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