Hi there, it is a rather old thread but I would like to draw your attention on a new paper that I found very interesting. Currently I am testing the method and would be nice to have some opinions on...

I have made hundreds of mutants using a modified SDM protocol. I have never used PAGE purified primers (for standard SDM- I have used them for ligation independent cloning). I always use the standard...

If the amino acids you want to mutate are so close together, and there are so many of the, you may find it easier to make your mutants by overlap extention PCR (aka "old-fashioned PCR mutagenesis")....