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:
>Hello all,
>  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.

Manuel

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