2 Rounds of PCR
jgraham at bronze.ucs.indiana.edu
Wed Mar 3 13:36:07 EST 1993
I'm sorry I can't respond to the question directly, however using two
rounds of PCR of 30-40 cycles is an excellent mutagenesis strategy, depending
on the quantities involved.
A discussion that passed here suggested that one round of 30 cycles might
lead to an efficient mutation frequency, roughly leading to an average of
about 4 base changes per several hundred under the conditions used.
We avoid this problem by limiting our amplifications to 16 rounds and working
with the bulk PCR product directly when possible.
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