Site directed mutagenesis kit

venita de almeida venita at merle.acns.nwu.edu
Tue Dec 20 10:50:04 EST 1994


In article <3d6mit$46p at delphi.bc.edu>, Thomas C. Chiles
<Chilest at HERMES.BC.edu> wrote:

> Can anyone direct us to a kit or method that will afford site directed
> mutagenesis of a gene without having to perform a lot of subcloning steps
> etc.  In short, we have subcloned a gene at great time and cost into a
> eukaryotic expression vector which affords expression of the gene product
> in our B cell "model" lines.  We would like to change several nucleotides
> within the gene while keeping it in the expresson vector.  From a novice
> standpoint, 1) what is the best approach (any references?); 2) is there a
> kit on the market that aid us in this endeavor?
> 
> Steve Amato, Chiles' laboratory
> ChilesT at hermes.bc.edu

you could use the USE mutagenesis kit from pharmacia .. or the kit from
clonetech ( i do not remember the name ) .. in these kits you use the
construct in whatever plasmid you have it in ... no subcloning at all 
the pharmacia kit is based on the paper by deng and Nickoloff ... Anal
Biochem 200 pp 81-88 ( 1992) 
Several of us  have used this and it has worked quite well 
if you have any more questions you can email me 
venita



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