Methods for developing TS mutant libraries in S. cerevisae
Manuel G. CLAROS
claros at biologie.ens.fr
Wed Jan 12 00:31:33 EST 1994
In Article <2gv183$3ic at hub.ucsb.edu>, upagan at mcl.ucsb.edu (Pa Gan) wrote:
> I'm currently working on developing a TS mutant library within a mutant
> strain of S. cerevisae, however, using my current methods, i am not
> generating the ammounts of TS mutants for my library.
> My current method is to perform mutagenesis on my strain with EMS, and
> grow them at 30C, then replica plate them and grow them at 37C and look
> for colonies which haven't grown. This method seems pretty straight
> forward however i have gotten very few TS mutants from my strain. Does
anyone know of a better method for developing a TS mutant library? Thanks much
> in advance for your replies! Pat.
> gan at lifesci.ucsb.edu
> upagan at mcl.mcl.ucsb.edu
Perhaps trying to isolate mutants at 23C (permisive temperature) you could
isolate TS colonies which can not grow at 30C.
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