QuikChange with Different plasmidsPCR mutagenesis
Posted 20 September 2012 - 11:26 AM
I have been able to get a mutation to work using one plasmid, but I got another plamsid with the same sequence (only the protein is tagged with an epitope for purification) and the PCR mutagenesis does not work.
Any thoughts as to why the same mutagenesis works on the same protein sequence (I have confirmed it's the same sequence) in one plasmid but not work in a differen plasmid?
Posted 20 September 2012 - 01:11 PM
Are you sure that the kit is OK right now? (if you still use the old one) (dNTPS aren't degraded, ...)
And what exactly means "does not work"? You got no colonies, you got only wt, you can't see a product after PCR mutagenesist?
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