yeast two hybrid (Y2H) libraries

[abble]

I'm hoping people will be willing to share their experiences on using yeast two hybrid libraries. There are a variety of options (e.g. commercial libraries, make your own, use libraries from another lab) and I'm wondering, as someone quite new to yeast two hybrid, what are some common pitfalls and things to avoid as well as where people have had success. If possible, please detail how the library was constructed, RNA source, complexity, etc.

[TheTheoretician]

I was at Stony Brook when Stan Fields developed it and I myself performed 3 successful two-hybrid screens. Back in the day, all libraries we made ourselves - with REs and molecular cloning - very poor genomic coverage. The last successful screen I did was with a sheared yeast genomic Clontech library (2001) which was stated to have 99% coverage and fragments ranging from 400 bp to 4.0 kb, give or take. I know I contacted Clontech about making a custom library, too.

 

Peace,

 

Ik

Edited August 17, 2010 by TheTheoretician