Site directed mutagenesis

Andy Phillips PHILLIPSA at UK.AC.AFRC.LARS
Tue Oct 27 03:47:00 EST 1992


Dear PCRologists and homebrewers,

I need to do some mutagenesis on a 500 bp stretch of DNA, to
create restriction sites about every 100 bp. I haven't done
any multiple site-directed mutagenesis since the dark ages B.P. 
(Before PCR) and haven't kept track of the methodology papers.
For cost reasons, I would prefer to make only one mutagenic
primer per site. Has anyone out there tried (with success) the
"megaprimer" method (Sarkar & Sommer BioTechniques (1990) 8:404-7)?
Or do you know a better method?

Thanks
Andy Phillips (PHILLIPSA at LARS.AFRC.AC.UK)
Lomg Ashton Research Stn, Bristol, UK



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