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[anonymous]

I have cloned a receptor which contains a single base insertion causing a frame shift mutation.  Due to time restraints and contamination problems it is not practicle to go back and redo PCRS.  

Im trying to delete the inserted base using a Clonetech kit where you introduce a mutation to the receptor and one to the expression vector (pcDNA3) for slection purposes:

1.  Has anyone used this method and if so does it have a high success rate?

2.  My mutation is ~500bp away from the site that I will mutate in the vector - is this ok?

3.  Any general advice about site directed mutagenesis in this setting?

[anonymous]

Have you thought about the Quickchange kit form Stratagene, its worked every time in our lab.