Best mutagenesis strategy? FAQs?

Julian Parkhill J.Parkhill at bham.ac.uk
Thu Nov 25 10:58:35 EST 1993


In article <CH1vyL.GMw at usenet.ucs.indiana.edu>,
jgraham at bronze.ucs.indiana.edu (the End) wrote:

> I have yet to find a mutagenesis technique that doesn't require sublconing
> the target either :)
> 
> Jim

Strange you should mention that, because just yesterday someone showed me a

paper that claims 58-97% mutagenesis with a single primer, no subcloning,
no special vectors, and no nucleotide analogues.

The reference is Lai et al, (1993) Nuc. Acids Res. Vol.21, No.7,
pp3977-3980

The method involves a very clever trick using 3 pre-existing restriction
sites (none of which are lost) and a varient of the gapped-duplex method.
I personally have not tried this (yet), but I have second-hand information
that it works as well as it claims.
-- 
Julian Parkhill [J.Parkhill at UK.AC.BHAM (JANET) or @BHAM.AC.UK (INTERNET)]
       CRC Laboratories, Dept. of Cancer Studies, Medical School,
     University of Birmingham, Edgbaston, Birmingham, B15 2TJ, U. K.
Disclaimer: If I thought anybody gave a damn about my opinions....



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