Abstract
Automated or semi-automated high-throughput RNAi screens are highly prone to systematic errors because of multistep repetitive protocols and extensive use of automated instruments. A well-designed RNAi library can help detect and minimize systematic human and robotic errors. In this unit, we describe how to design an RNAi bacterial library for use in conjunction with the well-studied nematode Caenorhabditis elegans for automated phenotypic screens. We provide strategies to design and assemble RNAi libraries to reduce or eliminate systematic errors. These strategies serve as a good quality-control check and facilitate obtaining high-quality data from a genome-wide and sub-library RNAi screen. © 2018 by John Wiley & Sons, Inc.