title: Next-Generation Sequencing for Identification of EMS-Induced Mutations in Caenorhabditis elegans.
abstract: 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.
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.|Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Nicolas J
authors.initials: NJ
authors.lastname: Lehrbach
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.|Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Fei
authors.initials: F
authors.lastname: Ji
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.|Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Sadreyev
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods; D004247:DNA / Q000235:genetics; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D016296:Mutagenesis / Q000187:drug effects; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation / Q000187:drug effects
chemical_list: D009153:Mutagens; D004247:DNA; D005020:Ethyl Methanesulfonate
keywords: C. elegans; EMS mutagenesis; next-generation sequencing