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  • Title: Identification of Protein-Protein Interactions Using Pool-Array-Based Yeast Two-Hybrid Screening.
    Author: Lathouwers T, Wagemans J, Lavigne R.
    Journal: Methods Mol Biol; 2018; 1794():29-48. PubMed ID: 29855949.
    Abstract:
    Array-based yeast two-hybrid (Y2H) screening in combination with a pooling strategy permits identification of complete interaction networks. After discussing the general advantages and drawbacks of Y2H, this chapter provides a detailed protocol for an array-based Gal4p Y2H screen using a pooling strategy to improve efficacy of the experimental work. This includes bait transformation, bait autoactivation testing, pooling of the baits and preys, yeast mating and evaluation of the results. Moreover, a one-on-one confirmation protocol and a quantitative α-galactosidase assay protocol to compare interaction strengths are provided.
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