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Yeast 2-Hybrid selection media - (Oct/27/2006 )

We are trying to confirm the protein-protein interaction of our candidate clones using SD drop out media minus Adenine, Histidine, Leucine, and Tryptophan. We just want to show that the yeast candidates don't grow if the interacting protiens aren't interacting. The main question I have is, is there a protocol that incorporates 3AT into the SD drop out media, or is it always used only on plates?

-molgennola-

QUOTE (molgennola @ Oct 27 2006, 09:07 AM)
We are trying to confirm the protein-protein interaction of our candidate clones using SD drop out media minus Adenine, Histidine, Leucine, and Tryptophan. We just want to show that the yeast candidates don't grow if the interacting protiens aren't interacting. The main question I have is, is there a protocol that incorporates 3AT into the SD drop out media, or is it always used only on plates?


I think most protocols use it only on plates, but you can (and we do in our lab) grow the clones in liquid dropout with 3AT......when we have a lot of clones, that's usually what we do (using 96 well plates).....

-shurgood-

do you have any sort of protocol for adding the 3AT to the liquid drop out?

-molgennola-