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High-quality yeast-2-hybrid interaction network mapping.

Curr. Protoc. Plant Biol. 26:e20067 (2018)
© 2018 John Wiley & Sons, Inc. In this article, we describe a Y2H interaction mapping protocol for systematically screening medium-to-large collections of open reading frames (ORFs) against each other. The protocol is well suited for analysis of focused networks, for modules of interest, assembling genome-scale interactome maps, and investigating host-microbe interactions. The four-step high-throughput protocol has a demonstrated low false-discovery rate that is essential for producing meaningful network maps. Following the assembly of yeast expression clones from an existing ORFeome collection, we describe in detail the four-step procedure that begins with the primary screen using minipools, followed by secondary verification of primary positives, identification of candidate interaction pairs by sequencing, and a final verification step using fresh inoculants. In addition to this core protocol, aspects of network mapping and quality control will be discussed briefly. © 2018 by John Wiley & Sons, Inc.
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Publication type Article: Journal article
Document type Scientific Article
Keywords High-throughput ; Interactome ; Network ; Protein-protein Interaction ; Systems Biology
ISSN (print) / ISBN 2379-8068
e-ISSN 2379-8068
Quellenangaben Volume: 26, Issue: 3, Pages: , Article Number: e20067 Supplement: ,
Publisher Wiley
Publishing Place Chichester
Reviewing status Peer reviewed
Institute(s) Institute of Network Biology (INET)