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random plasmid mutagenesis

David Rose drose at husc4.harvard.edu
Thu Aug 12 10:55:41 EST 1993


Hello:

	I am going to be doing a plasmid shuffle screen for mutants, and
am trying to decide what kind of mutagenesis to do.  The gene in question
has already been mutagenized with hydroxylamine, and I am interested in
other methods that make other basepair changes.  I had considered doing
PCR mutagenesis, but my gene has lousy sites and subcloning after
amplification would be a pain.  So, I would ideally like a method, like
hydroxylamine, in which one simply zaps the intact plasmid.  Is anyone
using any other methods, like different chemicals, passage through a
mutator bacteria strain, etc?  What works, what doesn't?  Any info would
be appreciated.

						Dave Rose
						drose at husc.harvard.edu



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