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-   -   Small yeast colonies on 2-hybrid screening plates (http://www.molecularstation.com/forum/yeast-forum/86430-small-yeast-colonies-2-hybrid-screening-plates.html)

Michele88 09-13-2012 12:26 PM

Small yeast colonies on 2-hybrid screening plates
 
Dear All,

I am doing some Y2H screens (with intrabody libraries) with the L40 strain (his3 -). I have a problem: when I plate onto selective -WHL + 3AT media, colonies do not grow well and remain small. What could it be? I tried with a positive control (a previously isolated intrabody binding the LexA DBD and the growth is very good onto same plates).
I am using a controlled bait line.

Your help is appreciated, thank you very much

Michele

pigfarmer 09-20-2012 11:12 PM

Re: Small yeast colonies on 2-hybrid screening plates
 
Quote:

Originally Posted by Michele88 (Post 442078)
Dear All,

I am doing some Y2H screens (with intrabody libraries) with the L40 strain (his3 -). I have a problem: when I plate onto selective -WHL + 3AT media, colonies do not grow well and remain small. What could it be? I tried with a positive control (a previously isolated intrabody binding the LexA DBD and the growth is very good onto same plates).
I am using a controlled bait line.

Your help is appreciated, thank you very much

Michele

How do youmean the colonies are small? Do the cells grow well prior to mating? Maybe there just isn't any interaction, or very slight?


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