Protocol

Two Hybrid Technologies

Volume 812 of the series Methods in Molecular Biology pp 21-38

Date:

Array-Based Yeast Two-Hybrid Screens: A Practical Guide

  • Roman HäuserAffiliated withKarlsruhe Institute of Technology
  • , Thorsten StellbergerAffiliated withKarlsruhe Institute of Technology
  • , Seesandra V. RajagopalaAffiliated withJ Craig Venter Institute (JCVI)
  • , Peter UetzAffiliated withCenter for the Study of Biological Complexity Virginia Commonwealth University Email author 

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Abstract

Yeast two-hybrid screens are carried out as random library screens or matrix-based screens. The latter have the advantage of being better controlled and thus typically give clearer results. In this chapter, we provide detailed protocols for matrix-based Y2H screens and give some helpful instructions how to plan a large-scale interaction screen. We also discuss strategies to identify or avoid false negatives and false positives.

Key words

ORFeome Mating Pooling Protein–protein interactions Yeast two hybrid Array Vectors Yeast strains