Invitro mutagenesis

Dr. Duncan Clark news at hgmp.mrc.ac.uk
Thu Mar 29 11:27:04 EST 2001


In article <v04011702b6e8f858d7e5@[144.92.64.179]>, the eminent Michael
L. Sullivan at BIOSCI/MRC Human Genome Mapping Project Resource Centre
wrote
>>I have been trying to do invitro mutagenesis using quickchange mutagenesis
>>kit
>>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.

Duncan 
-- 
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
http://www.dnamp.com
http://www.genesys.demon.co.uk




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