PCR

A PhD student of mine asked me about plasmid maps for several DNA constructs that I had created some 20 years ago.

If you need to modify your DNA construct, one frequently used method is the PCR-based mutagenesis, where you incorporate the mutation into the middle of two complementary primers.

My last NEBuilder assembly got stuck because I couldn't get two of the PCR reactions to work. So I tested alternatives to the Phusion DNA polymerase, which I have been using exclusively for cloning purposes during the last years...