site directed mutagenesis

Ashok Aiyar ashok at
Fri Aug 26 09:11:05 EST 1994

In article 6uF at,  ez001427 at () writes:
>Bulletin board login (maryr at 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% 
>	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.


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

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