Invitro mutagenesis

Dr. Duncan Clark news at
Thu Mar 29 11:27:04 EST 2001

In article <v04011702b6e8f858d7e5@[]>, the eminent Michael
L. Sullivan at BIOSCI/MRC Human Genome Mapping Project Resource Centre
>>I have been trying to do invitro mutagenesis using quickchange mutagenesis
>>from stratagene, somehow its not working with my plasmid, 

I have a post-doc who swears anything in pUC19 will not work with this
kit. So far for the one mutation we've tried recently here in a 700bp
insert in pUC19 failed miserably. As it was at the end of the insert we
eventually just used a long primer containing the mutant sequence and
used that in a straight PCR (with minimal cycles, Pfu and lots of
template) and generated it that way.

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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