Ethyl methanesulfonate (EMS) mutagenesis and DTT selection

AW Alan D. Winter
ET Elissa Tjahjono
LB Leonardo J. Beltrán
IJ Iain L. Johnstone
NB Neil J. Bulleid
AP Antony P. Page
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Synchronised L4 larvae were exposed to freshly diluted 50 mM EMS (Sigma M0880) for 4 h at room temperature (ref: “C. elegans: a practical approach”, chapter “Conventional Genetics” by Jonathan Hodgkin, “Protocol 1- EMS Mutagenesis”, pages 252-253) and selected for increased survival NGM-OP50-1 plates with on 5–7.5 mM DTT. A total of approximately 100,000 genomes were mutagenised in three separate screens with each screen being divided into sub-population to identify independent mutants.

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