Site Directed Mutagenesis Kits

Richard J. Dudley rdudley at nrc.uab.edu
Mon Jun 16 13:31:26 EST 1997


Vernon Wayne Bauer wrote:
> 
> I need to do some basic site-directed mutagenesis.  Nothing special.  It
> seems to me one of the commercial kits should be the easiest way, but I know
> nothing about any of them.  Does anyone have any thoughts, good/bad experiences,
> recommendations, etc. they could pass on?  Thanks for your help.
> 
> Vernon Bauer
> Genetics/NCSU

We use Promega's Altered Sites kit and always obtain good results.  You
have to use the single stranded DNA protocol, though--us and several
other labs haven't had much success with the "new" method.  This kit
requires youy to clone into pAlter, but I think they're coming out with
one that doesn't require additional subcloning.

We get between 50-90% mutagenesis about 98% of the time.  We've made
maybe 50 mutants this year alone, and only one didn't mutate the first
try (second try worked, though).  Very rarely fails us.

(no affiliation, just a happy customer).

--- --- ---
Richard J. Dudley (rdudley at nrc.uab.edu)                            
Department of Neurobiology                    
University of Alabama School of Medicine       
http://www.nrc.uab.edu/



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