Site-directed mutagenesis

Michael Szardenings msz at bmc.uu.se
Tue Jun 4 08:19:18 EST 1996


In article <4oepg1$t9b at ratatosk.uio.no>, eijohnse at bioslave.uio.no says...
>
>I am embarking on some site-directed mutagenesis and I see that Stratagene
>offers two different types of kit: One which is PCR based (ExSite) and one
>which uses T7 polymerase (Chameleon).  Has anyone had any experience with
>either of these systems and if so what would you advise? Any advice on
>other systems would also be welcome.

Dear Oyvind,
if you have an average mol-biol lab, you don't need to buy kits from 
companies. With a very few exceptions most kits are based on some 
publications which give good descriptions of the technology and few special 
strains and/or enzymes must be purchased, but most will already be in your lab 
or can easily be obtained by contacting other labs.
Personally I have been sticking to PCR based mutagenesis in the last years. 
It usually requires nothing above what you already may have (vector, 
sequencing primers, PCR equipment) and a primer to carry out the mutagenesis.
There is a whole bunch of papers published about different variants. If you 
can, check MEDLINE or similiar for papers in journals present in your library. 
Some methods have been reinvented 4-5 times, but you might find something that 
fits to your special purpose.

Good luck		Michael
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