PCR

ckf craig at fulton52.freeserve.co.uk
Thu Dec 9 10:25:56 EST 1999


Does anyone have a decent method of calculating the annealing temp. for
degenerate PCR primers? I've been using primer premier  but I'm not sure if
the values are reliable. The primers also contain inosine and I don't think
that the program compensates for that.

On a related theme, I've been doing colony PCR on a few different types of
bacteria and I generally find that if the reaction fails there's a hint of
chromosomal DNA on my gels. I would assume this means too many cells in the
reaction. I was wondering what sort of things can be released on lysis that
will inhibit the reaction so much.

Cheers

Craig





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