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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