Next-Generation Sequencing for Identification of EMS-Induced Mutations in Caenorhabditis elegans

• Published in: Current protocols in molecular biology (Print) Vol. 117; p. 7.29.1
• Main Authors: Lehrbach, Nicolas J, Ji, Fei, Sadreyev, Ruslan
• Format: Journal Article
• Language: English
• Published: United States 05.01.2017
• Subjects: Animals; Caenorhabditis elegans - drug effects; Caenorhabditis elegans - genetics; Chromosome Mapping - methods; DNA - genetics; Ethyl Methanesulfonate - pharmacology; High-Throughput Nucleotide Sequencing - methods; Mutagenesis - drug effects; Mutagens - pharmacology; Mutation - drug effects; next-generation sequencing; EMS mutagenesis; C. elegans
• ISSN: 1934-3647
• DOI: 10.1002/cpmb.27

Forward genetic analysis using chemical mutagenesis in model organisms is a powerful tool for investigation of molecular mechanisms in biological systems. In the nematode, Caenorhabditis elegans, mutagenesis screens using ethyl methanesulfonate (EMS) have led to important insights into genetic control of animal development and physiology. A major bottleneck to this approach is identification of the causative mutation underlying a phenotype of interest. In the past, this has required time-consuming genetic mapping experiments. More recently, next-generation sequencing technologies have allowed development of new methods for rapid mapping and identification of EMS-induced lesions. In this unit we describe a protocol to map and identify EMS-induced mutations in C. elegans. © 2017 by John Wiley & Sons, Inc.