E. coli mutator strains

Dan Zabetakis dan at cubsps.bio.columbia.edu
Fri Jul 7 13:56:08 EST 1995


In article <3tgsc9$p9r at riscsm.scripps.edu>,
SLForsburg  <susan_forsburg at qm.salk.edu> wrote:
>
>Stratagene just sent me a flyer about XL1-Red (I have no affiliation 
>with them).  The strain has *three* different mut mutations and must be 
>unstable as all get-out, but they claim a high level of plasmid hits 
>just in the process of forming colonies on plates.  Sounds like a great 
>idea to me and much much nicer than PCR or hydroxylamine.  They are 
>selling competent cells.  I dont know how much they cost.

  I and one other person in my lab used them, but without success. That's
probably more on the nature of our genes as this was not the first attempt
to get mutants. We were looking for ts mutants in our respective genes.
Mine was not essential and my lab mate's was.
  The benefits in theory are that the mutagenesis is easy and quick, and 
hits every base pair rather than specific ones like chemical treatments.

  It is quick. Just transform your plasmid, collect the transformants, 
grow for whatever time you think gives the right amount of mutagenesis,
do a big prep. Then you can transform into yeast and look for the effect
you want. I was looking for a ts mutant. I didn't get it but that might be
because my gene isn't essential and there just isn't a ts version to be
had.
  It seems like a good system but I can't really endorse it because I
didn't get what I wanted.

DanZ

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