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Updated: May 30, 2026

Isolating Interaction-Null/Impaired Mutants Using the Yeast Two-Hybrid Assay
Published on: December 29, 2023
Media composition influences yeast one- and two-hybrid results
Ying Liu1, Zabeena Merchant1, Hao-Ching Hsiao2
1Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005 USA.
Abstract:
Although yeast two-hybrid experiments are commonly used to identify protein interactions, the frequent occurrence of false negatives and false positives hampers data interpretation. Using both yeast one-hybrid and two-hybrid experiments, we have identified potential sources of these problems: the media preparation protocol and the source of the yeast nitrogen base may not only impact signal range but also effect whether a result appears positive or negative. While altering media preparation may optimize signal differences for individual experiments, media preparation must be reported in detail to replicate studies and accurately compare results from different experiments.

