site directed mutagenesis using synthesized oligos - (Oct/17/2007 )
I need to make 3 amino acid changes in a gene sequence. Two of them are pretty close to each other (positions 87 and 96) but the third one is far off (position 244). I am thinking of ordering synthesized oligos with the mutations incorporated and with restriction sites incorporated at the ends. Then I can cut the wild type sequence with the restriction enzymes and simply ligate the synthesized oligos and get my mutations incorporated into the gene this way. Has anyone tried this approach before? Is this feasible? Please help.