DNA point mutation - Hard to mutate DNA which has GC content high (Nov/23/2004 )
I am just doing the DNA point mutation using Stratagene's pfuUltro and Turbo and follow the manual of stratagene Quickchange kit. I can get clonies but the DNA were not mutated or DNA were wild type. I tried many times with differnt conditions. But ,. After we discussed, we think the DNA has high GC content and dificult to do mutation because PCR doesn't work. Did you meet the similiar situation? Could you give some sugestion?
Our lab has a reliable source for "custom gene synthesis" when dealing with GC-rich, hard-to-amplify, hard-to-clone low-copy sequences (check their website on www.epochbiolabs.com). They can synthesize any sequence, any size and 100% sequence quaranteed in a short period of time. Just compare the time, cost and your purpose, custom gene synthesis could be a way to go.