Mapping random mutagenesis

bbraun at ucla.edu bbraun at ucla.edu
Tue Feb 20 05:18:00 EST 1996


I am thinking of a project that would entail random mutagenesis of
a 300-1500 bp promoter fragment.  My question is, is there any way
to identify where a mutation lies to within a few base pairs short
of sequencing the construct?  That way I could assess an entire
population of mutants in one reaction - to assess the randomness
of mutation, and after a genetic selection to look for clusters of
mutations.  For example, I was considering using RNase protection
against a library of mutants.  In an ideal mutagenesis I would get
cleavage at each base with equal frequency, but hot spots would
show greater cleavage frequency, and therefore darker bands.
Linker insertion would be great, but I don't know of an easy way 
to get good distribution of the linkers.

Any ideas out there?  I'd love to hear some!

Thanks,

Benjamin Braun
bbraun at ucla.edu



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