Small yeast colonies on 2-hybrid screening plates

[Michele88]

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]

Originally Posted by Michele88

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?