random Mutagenesis

Daniel Kim dkim at nmsu.edu
Thu Jul 20 14:05:51 EST 1995


In article <3ujpne$1r9 at news.wesleyan.edu> Visu <rvisvanathan at eagle.wesleyan.edu> writes:
>Hi,
>
>I need to do random mutagenesis on a 3kb fragment.
>Is there any efficient way of doing the mutation,
>say by PCR?
>
>If you know of any reference or protocol, please
>let me know.
(omitted stuff)

There is a method called DNA shuffling mutagenesis by a guy named Willem 
Stemmer, published in SCIENCE and/or PNAS in late 1994/early 1995.  He 
takes a PCR fragment, DNAse I digests it and purifies out 50 - 200 bp 
fragments, then run PCR cycles with NO primers to let the fragments 
anneal and primer off each other, introducing recombination errors as 
well as PCR misincorporation errors.


Sorry, no specific citation, you'll have to look for it.  :(

Daniel Kim




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