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Axenic and Synchronous Cultures of Caenorhabditis Elegans

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For more content, see Nematology.

A technique to axenize cultures of Caenorhabditis elegans on a large scale is described. This procedure has enabled the isolation of eggs of C. elegans in quantities sufficient to perform biochemical tests. Adults from monoxenic cultures were used to isolate eggs. When egg-bearing worms were suspended for 2-3 hr at 25o in an alkaline (0.4 M) solution of NaOH, the worm cuticle was partially digested, releasing free eggs. The high alkalinity rendered the eggs bacteria-free. Eggs were then isolated from a linear sucrose gradient. Eggs from the least dense band (1.13 g/cm3) hatched within 10-13 hr when resuspended in a liquid medium and yielded a synchronous culture. The hatchability of these eggs ranged from 20-35%. Inhibitory effects of 5-fluorodeoxyuridine and hydroxyurea on freshly hatched (L1 stage) worms are described. Levels of isocitrate lyase were determined in eggs and in synchronous cultures.

Affiliations: 1: Program in Biochemistry and Biophysics, Chemistry Department, Washington State University, Pullman, Washington 99164, U.S.A.


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