PCR mutagenesis

Steve Minchin S.D.Minchin at bham.ac.uk
Mon Jun 20 09:44:45 EST 1994

In article <2tsoar$n17 at masala.cc.uh.edu> bchs1b at Elroy.UH.EDU (Michael Benedik) writes:
>From: bchs1b at Elroy.UH.EDU (Michael Benedik)
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: PCR mutagenesis
>Date: 17 Jun 1994 17:59:55 GMT
>Organization: University of Houston
>Lines: 12
>Message-ID: <2tsoar$n17 at masala.cc.uh.edu>
>Reply-To: bchs1b at Elroy.UH.EDU
>NNTP-Posting-Host: elroy.uh.edu

>Does someone have the protocol/reference for doing random mutagenesis
>by PCR amplification of a target fragment. I think you just change
>nucleotide concentrations and/or add Mn++, but I don't have the details.


> Michael Benedik                                INTERNET: Benedik at uh.edu
> Dept. of Biochemical & Biophysical Sciences    
> University of Houston                          BITNET: Benedik at uhou

Hi Michael,

I answered a similar query about a week ago. We have used PCR mutagenesis 
quite successfully. We do have a good selection for the desired mutants and 
therefore we normally screen many thousands of colonies.

It is not possible to use just one protocol for mutagenesis of different 

There is a good review of PCR conditions and the fidelity of DNA polymerases 
by Eckert and Kunkle in the book:

PCR - A Practical Approach, Edited by McPherson, Quirke and Taylor.
Published by IRL Press at Oxford University Press.

I hope this helps.

All the Best


Dr. Steve Minchin,

School of Biochemistry, The University of Birmingham,
Edgbaston, Birmingham, B15 2TT, U.K.

InterNet:       S.D.Minchin at bham.ac.uk
Janet:          S.D.Minchin at uk.ac.bham


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