Polymerase for Quikchange mutagenesis - (Nov/21/2007 )
Hi, I am trying to do site directed mutagenesis based on teh Quikchange principle but without actually buying the kit. The kit comes with Pfuturbo polymerase for the PCR reaction. I don't have that enzyme and don't want to buy it. Can I use Phusion or pwo instead? Has anyone tried this? Do these enzymes have strand displacing activity?
There's a mutagenesis kit for Phusion also, so I think you can use it in the exact same way as you would Pfuturbo. Don't know about pwo, but isn't that one also isolated from a pyrococcus species (as pfu orignially also was)? (maybe they have rathers simillar properties then).
You could use it with phusion. You might need to adjust the extension time.
Well, you need a high fidelity polimerase and remember Pfu don’t add A and the end.