Degenerate PCR: concentration of primers?

Markus Piotrowski Markus.Piotrowski at ruhr-uni-bochum.de
Tue Nov 4 16:25:52 EST 1997


Dear Huang,

I had success with PCR using degenerate primers. The PCR-cocktail was 'normal',
that means a final volume of 50 µl and a dNTP concentration of  0.2 mM each. Our
primers were about 512x degenerated and in some cases, to avoid an N we used
inosine. The primer concentration was high: 400 pmol of each primer and we used
touch-down PCR. This worked for us with cDNA as well as with a cDNA-library. I
hope this will help you,

Markus







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