site directed mutagenesis

[Ashok Aiyar]

In article 6uF at ucdavis.edu,  ez001427 at dale.ucdavis.edu () writes:
>Bulletin board login (maryr at magnus1.com) wrote:
>: I anticipate doing some site directed mutagenesis in the near future.  I would
>: like some recommendations from folks on which mutagenesis kits you have used
>: and the pros and cons of them. Thanks, Mary
>
>	I've used the Amersham site directed kit extensively, and found 
>it to be excellent.  Also, the kit is such that you buy it once, and just 
>buy needed enzymes and such as they run out.  The kit is expensive, but 
>it's much better than the muta-gen type kits (dut ung).  Around 90% 
>mutants.
>
>	I like the idea of response #1, the PCR method, bu most of those 
>that I've seen require two primers per mutation site, which can get 
>pricey if you have 50 or 100 specific mutants to create.
>

I have used PCR mutagenesis extensively, with one mutant oligo per
mutation.  In addition, two external primers are required, which 
flank convenient restriction sites.

Later,
Ashok


--
Ashok Aiyar                              email: ashok at mirage.bioc.cwru.edu 
CWRU Medical School                              telephone: (216) 368-3300 
Department of Biochemistry                             fax: (216) 368-4544