Can anyone give me a technical details (steps involved) on how to do PCR mutagenesis using four primers.
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1 reply to this topic
Posted 16 July 2001 - 09:00 PM
You do 2 individual PCr with 2 sets of primers, the mutated base in contained with in 2 of the primers, one of each used in the initial PCR. These primers overlap by at least 10 bases.You then do a second PCR, using the flanking primers ie those which do not contian the mutation using your 2 initial PCR products as a template, this should give you a product containing the mutation.