U.S.E. Mutagenesis Kit

Curt Ashendel ashendel at aclcb.purdue.edu
Tue Jan 28 12:16:09 EST 1997


On Mon, 27 Jan 1997 18:36:33 +0100, Stefano Santanche'   
<sast at axrma.uniroma1.it> wrote:

>I need some informations about the U.S.E. Mutagenesis Kit. 
>a) did anyone performe double mutation in a single round of mutagenesis,
>using two mutagenic oligos and the selection one at the same time?
We have not tried this, so I can't say.

>b)Is it possible to make the primary restriction enzyme selection taking
>advantage of the fact that the mutagenic primer  eliminates a
>restriction site?.

Yes. We do this all the time. So far (a dozen or two mutants) I have 
always been able to design the mutagenesis oligo to either create or 
delete a  mappable restriction site so we can screen by mapping. It 
does take a lot a staring at the sequence possibilities to find short 
palindromes. It is easier if you have easy access to unusual 
restriction enzymes, but so far I have been able to keep to the common 
10 enzymes for all but one mutation.

y

Curt Ashendel
Purdue University, West Lafayette, IN
ashendel at purdue.edu



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