QUIK change - (Oct/11/2007 )
I'm confused as to how quik change works. Is it the same as doing PCRs? Also, what is the advantag of doing quik chang over doing PCR for insertion or deletions?
Quikchange amplifies the plasmid aswell as your gene so doesn't require any further sub-cloning. You just need to screen then sequence minipreps. try the manual for details.
Quick change amplifies the entire plasmid using primers you specifiy and ~80% of the colonies you screen will have the mutation you want. I've had really good luck whenever I use the quick change kit. You MUST follow the directions EXACTLY, the right falcon tubes and everything. The other way has MUCH lower efficency. You can find the manual here....
Thanks for the replies. So QUIK change does not amplify the plasmid. With the QUIK change method, does anyone know what is the max number of sequences I can delete or insert?
I heard sb got 12bps insertion here.
I don't do anymore than 9 Nt.