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Direct cell to cell Yeast-E.coli shuttle by electroporation?

Nick Kaplinsky kaplinsk at ohsu.edu
Tue Aug 22 15:50:28 EST 1995


You may want to try using a Promega miniprep column (~$1) to purify your 
yeast plasmid.  The resin specifically binds plasmid DNA and whatever 
the yeast factor is that inhibits transformation gets washed away.  I 
even use the included alkaline lysis reagents on my yeast (no need for 
zymolyase or glass beads/phenol/chloroform) - I get 50-100 colonies by 
transforming with 5ul of the DNA I get from the prep this way (from a 
2ml liquid culture).  I am using this for screening two-hybrid clones, 
and by plating my E. Coli onto Leu- plates I do my curing in the same 
step as the tranformation.
Nick





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