yeast->E.coli electroporation

[Karlheinz R. Skowronek]

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 cytoplasm
to the E.coli (cell wall or cytoplasm?).  Has anyone used this protocol or
one like it with the BioRad gene pulser? 


Karlheinz


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