Yeast Two Hybrid Problem

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scottrik
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Yeast Two Hybrid Problem

Post by scottrik » Mon May 28, 2007 1:00 pm

I'm having some trouble with my Clontech yeast two hybrid construction and screening kit. I've run two assays so far as well as a positive and negative control. My second assay has just begun growth and it looks just like my first--->thousands of colonies. My negative control appears to be doing the same! Has anyone experienced this or does anyone have suggestions? I'm using defined medium from the manufacturer so I am almost positive the Quadruple dropout media I am using is satisfactory (-Ade, His, Leu, Trp). The fact that I've tried two different baits as well as controls (with the same results) indicates that autoactivation of the bait is unlikely (as far as I know). Any help anyone can give me would be greatly appreciated! Thanks!

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