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

doi:10.1002/cpmb.27

. 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

Nicolas J Lehrbach^{1

2}, Fei Ji^{1

2}, Ruslan Sadreyev^{1

3}

Affiliations

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.

PMID: 28060408
PMCID: PMC5303615
DOI: 10.1002/cpmb.27

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

Nicolas J Lehrbach et al. Curr Protoc Mol Biol. 2017.

. 2017 Jan 5:117:7.29.1-7.29.12.

doi: 10.1002/cpmb.27.

Authors

Nicolas J Lehrbach^{1

2}, Fei Ji^{1

2}, Ruslan Sadreyev^{1

3}

Affiliations

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.

PMID: 28060408
PMCID: PMC5303615
DOI: 10.1002/cpmb.27

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.

Keywords: C. elegans; EMS mutagenesis; next-generation sequencing.

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References

1. Afgan E, van den Baker D, Beek M, Blankenberg D, Bouvier D, Čech M, Chilton J, Clements D, Coraor N, Eberhard C, et al. The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update. Nucleic Acids Res gkw343 2016 - PMC - PubMed
1. Chu JSC, Chua SY, Wong K, Davison A, Johnsen R, Baillie DL, Rose AM. High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans. BMC Genomics. 2014;15:361. - PMC - PubMed
1. Davis M, Hammarlund M, Harrach T, Hullett P, Olsen S, Jorgensen E. Rapid single nucleotide polymorphism mapping in C. elegans. BMC Genomics. 2005;6:118. - PMC - PubMed
1. Doitsidou M, Poole RJ, Sarin S, Bigelow H, Hobert O. C. elegans mutant identification with a one-step whole-genome-sequencing and SNP mapping strategy. PLoS ONE. 2010;5:e15435. - PMC - PubMed
1. Fay D. Genetic mapping and manipulation: Chapter 1-Introduction and basics. WormBook. 2006 http://www.wormbook.org/chapters/www_introandbasics/introandbasics.html. - PMC - PubMed

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[1] Afgan E, van den Baker D, Beek M, Blankenberg D, Bouvier D, Čech M, Chilton J, Clements D, Coraor N, Eberhard C, et al. The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update. Nucleic Acids Res gkw343 2016 - PMC - PubMed

[2] Afgan E, van den Baker D, Beek M, Blankenberg D, Bouvier D, Čech M, Chilton J, Clements D, Coraor N, Eberhard C, et al. The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update. Nucleic Acids Res gkw343 2016 - PMC - PubMed

[3] Chu JSC, Chua SY, Wong K, Davison A, Johnsen R, Baillie DL, Rose AM. High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans. BMC Genomics. 2014;15:361. - PMC - PubMed

[4] Chu JSC, Chua SY, Wong K, Davison A, Johnsen R, Baillie DL, Rose AM. High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans. BMC Genomics. 2014;15:361. - PMC - PubMed

[5] Davis M, Hammarlund M, Harrach T, Hullett P, Olsen S, Jorgensen E. Rapid single nucleotide polymorphism mapping in C. elegans. BMC Genomics. 2005;6:118. - PMC - PubMed

[6] Davis M, Hammarlund M, Harrach T, Hullett P, Olsen S, Jorgensen E. Rapid single nucleotide polymorphism mapping in C. elegans. BMC Genomics. 2005;6:118. - PMC - PubMed

[7] Doitsidou M, Poole RJ, Sarin S, Bigelow H, Hobert O. C. elegans mutant identification with a one-step whole-genome-sequencing and SNP mapping strategy. PLoS ONE. 2010;5:e15435. - PMC - PubMed

[8] Doitsidou M, Poole RJ, Sarin S, Bigelow H, Hobert O. C. elegans mutant identification with a one-step whole-genome-sequencing and SNP mapping strategy. PLoS ONE. 2010;5:e15435. - PMC - PubMed

[9] Fay D. Genetic mapping and manipulation: Chapter 1-Introduction and basics. WormBook. 2006 http://www.wormbook.org/chapters/www_introandbasics/introandbasics.html. - PMC - PubMed

[10] Fay D. Genetic mapping and manipulation: Chapter 1-Introduction and basics. WormBook. 2006 http://www.wormbook.org/chapters/www_introandbasics/introandbasics.html. - PMC - PubMed

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

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

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