Yeast two-hybrid results - (Jan/23/2008 )

Hi everyone! I don't know if this is the right topic to post under but since Y2H tests for protein-protein interaction I'm going to give it a try. So I did a Y2H library screen, testing for the activation of the reporter genes B-gal and Uracil. However, I have gotten some conflicting results. When the yeast grows on the -uracil selective plates they don't turn blue on x-gal, but when the yeast turns blue on x-gal they don't grow on -ura selective plates . I have included a positive control on the selective plate and x-gal plate and both worked nicely, so I am sure the plates are good. Has anyone else encountered that? Please help!

Y2H is pretty tricky.

You always require replicates and repeats of your experiments. Only strong and reproducible results accouonts. In this case, you still need other approaches to further confirm your results.

Just discard these data with weak interactions, inconsistent results, most of which are false positive.

If you are still not sure, drop me a message. I can tell you a long story about 5 year experience with Y2H.

Hi everyone! I don't know if this is the right topic to post under but since Y2H tests for protein-protein interaction I'm going to give it a try. So I did a Y2H library screen, testing for the activation of the reporter genes B-gal and Uracil. However, I have gotten some conflicting results. When the yeast grows on the -uracil selective plates they don't turn blue on x-gal, but when the yeast turns blue on x-gal they don't grow on -ura selective plates . I have included a positive control on the selective plate and x-gal plate and both worked nicely, so I am sure the plates are good. Has anyone else encountered that? Please help!