Deletions by site directed mutagenesis

BSS032 at BANGOR.AC.UK BSS032 at BANGOR.AC.UK
Tue Mar 22 10:46:46 EST 1994


Rgearding site directed mutagenesis, ZURLINDEN at ch.unibe.IMB asks about the
size of deletions which can be made.

I have published data using the phosphorothioate method (Amersham supplya kit) in which we made deletions of 48 and 91 base pairs with high efficiency, i.e.
over 65%. I have also done deletions of a couple of hundred base pairs with
similar results. Primers are typically 30 mers.

See Sayers, Krekel and Eckstein, Rapid high efficiency site-directed mutagenesis by the phosphorothioate approach. BIOTECHNIQUES, Vol. 13, N0. 4, 592-596 (1992).

I should declare an interst, I am Sayers and get about 50 cents for each Amersham kit sold. 

See also Sayers and Eckstein Chapter 3 in Directed Mutagenesis-A practical
approach IRL Press (Ed. McPherson, M), 1991, pp49-69.
Olsen, Sayers and Eckstein, METHODS IN ENZYMOLOGY, (1993) Vol. 217, pp179-182.

Cheers, Jon Sayers,

Jon R Sayers (Cell and Molecular Biology), Biochemistry Dept.,
University College North Wales, Univ of Wales, Bangor, LL57 2UW.
Tel +44 248 38 23 54
Fax +44 248 370731
Email bss032 at uk.ac.bangor
or bss032 at bangor.ac.uk



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