2 Rounds of PCR

the End 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.

J. Graham

More information about the Methods mailing list