chemical mutagenesis

Mark Slack mt at
Thu Aug 8 15:14:17 EST 1996

I am currently trying to make a series of mutants by chemical mutagenesis in a 
300bp DNA fragment and I am aiming for single hit.  The method I am 
currently using is to incubate the DNA for 5mins in 0.7M NaNO2( in 1M NaAc 
PH 4.6). Subsequent sequencing of the clones produced showed no mutations in 
this area. I would appreciate it if anyone could supply me with a decent 
protocol for this, particularly regarding the amount of DNA to be used, the 
concentration of nitrous acid and the incubation times necessary.

Many thanks. 

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