Optimization of ENU mutagenesis of Caenorhabditis elegans.
Chemical mutagenesis of Caenorhabditis elegans has relied primarily on EMS to produce missense mutations. The drawback of EMS mutagenesis is that the molecular lesions are primarily G/C --> A/T transitions. ENU has been shown to produce a different spectrum of mutations, but its greater toxicity to C. elegans makes it a difficult mutagen to use. We describe here methods for minimizing ENU toxicity in C. elegans. Methods include preparing ENU stocks in absolute ethanol and storing stock solutions for not more than 2 weeks at -20 degrees C. To maintain reasonable brood sizes of mutagenized animals, mutagenic solutions should not exceed 1.0mM ENU. We provide data which suggest ENU is degraded or altered to more toxic products in aqueous solution, but less so in solvents such as absolute ethanol.
De Stasio
E A
EA
Department of Biology, Lawrence University, Appleton, WI 54911, USA. elizabeth.a.destasio@lawrence.edu
Dorman
S
S
D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004356:Drug Storage; D005038:Ethylnitrosourea / Q000737:chemistry / Q000633:toxicity*; D006863:Hydrogen-Ion Concentration; D016296:Mutagenesis / Q000187:drug effects* / Q000235:genetics; D009152:Mutagenicity Tests; D009153:Mutagens / Q000737:chemistry / Q000633:toxicity*; D020125:Mutation, Missense / Q000187:drug effects; D017354:Point Mutation / Q000187:drug effects; D012997:Solvents / Q000737:chemistry; D013997:Time Factors
D009153:Mutagens; D012997:Solvents; D005038:Ethylnitrosourea