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Curr Protoc Mol Biol. 2017 Jan 5;117:7.29.1-7.29.12. doi: 10.1002/cpmb.27.

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

Author information

1
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.
2
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
3
Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts.

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.

KEYWORDS:

C. elegans; EMS mutagenesis; next-generation sequencing

PMID:
28060408
PMCID:
PMC5303615
DOI:
10.1002/cpmb.27
[Indexed for MEDLINE]
Free PMC Article

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