Chemical Mutagenesis of DNA

[Jerry Salem]

Hello Netters,

Happy New Year!

We are trying to make random mutations in a plasmid.  Something on the
order of one random mutation event per plasmid.  We have had some marginal
luck using UV and EMS (methane-sulfonic acid ethyl ester).  

However, we were wondering if anyone has any experience with doing
chemical mutagenesis using these or other chemical mutagens (for instance
nitrosoguanidine) who would be willing to share some expertise.  As to
which methods give the highest number of random mutations and what
chemicals are easier to use.  We are particularly interested in AT
transversions although we also require AT to GC transitions.

Please respond via email since I don't get to this group very offen, I
will summarize all the responses and post them.

TIA


Jerry Salem

jerry at slack.med.upenn.edu