Hi Matthias Thanks for the info about self-SLIC. You wrote "This is because the T4 DNA polymerase treatment removes the newly synthesized strand in your mutated region (wich is your SLIC homology region) to have two 5' overhangs for annealing." Rather than "newly" don't you mean existing or template?
How long are the primers? What would the XX region be if one was making a single codon change? From the figure it looks like the forward strand is longer than the reverse strand. Was that intentional? And what are the advantages over a QuikChange type of reaction? Shorter primers, and more complete duplex plasmids? The amplification does not appear to be exponential. Many thanks for clarifying these points. -Mark Saper
