Chaudhuri, Jyotiska, Parihar, Manish and Pires-da Silva, André Francisco (2011) An introduction to Worm Lab : from culturing worms to mutagenesis. Journal of Visualized Experiments, Vol.47 . Article no. e2293. doi:10.3791/2293 ISSN 1940-087X.

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Abstract

This protocol describes procedures to maintain nematodes in the laboratory and how to mutagenize them using two alternative methods: ethyl methane sulfonate (EMS) and 4, 5', 8-trimethylpsoralen combined with ultraviolet light (TMP/UV). Nematodes are powerful biological systems for genetics studies because of their simple body plan and mating system, which is composed of self-fertilizing hermaphrodites and males that can generate hundreds of progeny per animal. Nematodes are maintained in agar plates containing a lawn of bacteria and can be easily transferred from one plate to another using a pick. EMS is an alkylating agent commonly used to induce point mutations and small deletions, while TMP/UV mainly induces deletions. Depending on the species of nematode being used, concentrations of EMS and TMP will have to be optimized. To isolate recessive mutations of the nematode Pristionchus pacificus, animals of the F2 generation were visually screened for phenotypes. To illustrate these methods, we mutagenized worms and looked for Uncoordinated (Unc), Dumpy (Dpy) and Transformer (Tra) mutants.

Item Type:	Journal Article
Divisions:	Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Journal or Publication Title:	Journal of Visualized Experiments
Publisher:	JoVE
ISSN:	1940-087X
Official Date:	2011
Dates:	Date Event 2011 Published
Volume:	Vol.47
Page Range:	Article no. e2293
DOI:	10.3791/2293
Status:	Peer Reviewed
Publication Status:	Published

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