Deng/Nickoloff mutagenesis

Scott Vande Pol sbv at
Mon Aug 28 16:46:48 EST 1995

We have been making site specific mutations by the Deng/Nickoloff technique
(aka clontechs "altered sites" kit or stratagenes "chameleon" kit) .  We
have had some undesired frameshift mutations at the ends of the
mutagenizing oligos, but have not yet found second site undesired
mutations.  Although T4 polymerase has a very low polymerase error rate,
other factors (methylation of the nucleotides, passage through  mutS
bacterial strains, etc ) may effect second site mutation rates.  Does
anyone know of any data ( or have personal experience) on the frequency of
undesired second site mutations using this procedure? 
	Thanks for any info on this!

Scott Vande Pol
Case Western Reserve University
Department of Pathology
Cleveland Oh. 44106
sbv at 

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