5-FOA selection

Dave Lloyd djl17 at leicester.ac.uk
Fri Aug 18 11:14:51 EST 2000


I have conducted a two hybrid screen and have a selection of positives. 
Bait plasmid's marker is URA3, and Library plasmid's marker is LEU.  Now i 
want to lose the bait plasmid to plasmid rescue the library plasmid, and 
also to check that this plasmid is not self-activating.  I pick a colony 
and grow in SD-LEU media till saturation then streak out on SD-Leu + 
0.1%FOA plates, but get nothing back, why is this??
I am attempting to do two rounds of culturin before i streak out on the FOA 
plates.  Or i could spin down my cultures and resuspend in a small volume 
and plate it all out on FOA plates however this would mean having one FOA 
plate per culture (or Y2H positive) thus a lot of plates, and lots of 
money....

************************
Dave Lloyd
Division of Medical Genetics
University of Leicester
Leicester
LE1 7RH
UK
Tel: +44 116 252 5088
Fax: +44 116 252 3378
************************


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