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yeast->E.coli electroporation

Karlheinz R. Skowronek kskowron at gnu.uvm.edu
Fri Jan 17 14:59:54 EST 1997




Hi sorry about the first post.  I don't know what happened.

Hi folks,

I'm trying to use an electroporation method to rescue plasmids from yeast
directly to E. coli.  I read a protocol in a supplement to the Red Book
(supl. 35 summer 1996) and a paper in Nucl.Acid Res. 1992 20:4.  The
protocol calls for mixing the the yeast and E.coli then hitting the cells
twice with varying amounts of voltage/current. The first electroporation I
assume is pushing the plasmid from the yeast nucleus to the cytoplasm and
the second electroporation is pushing the plasmid from the yeast cytopla
sm to the E.coli (cell wall or cytoplasm?). We've tried the procedure
unsuccessfully.  Has anyone used this protocol or one like it? 


Karlheinz

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