Worm Breeder's Gazette 2(1): 6
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
Our laboratory has been using two methods for mass growing of worms in preference to the liquid culture method of Sulston and Brenner. These methods are as follows 1. Growth on NGM plates with frozen E. coli.Two to four grams of frozen E. coli B cells (late log) are spread over the surface of a 100 mm NGM plate containing 30 ml of NGM media. These plates are innoculated with 20-50mg of young worms. (10+E4 to 10+E5 L1's may be obtained from over crowded N-2 stock plates). These plates are left for 2 to 4 days and then harvested by washing with M-9 buffer and cleaned by floatation in 35% sucrose. Single plate yields vary from . 4 gm to 1 gm per plate. Care must be taken not to leave worms on these plates too long since toxic products seem to accumulate past 4 days at room temperature. 2. Egg media on NGM plates. Take one egg; break into 100 ml beaker containing 50 ml of boiling water on a magnetic stirrer set at mid speed. Excess liquid is decanted off and the residue homogenized in an omnimizer at top speed for two minutes. This slurry is poured, 5 ml per plate, on top of standard 100 mm NGM plates. When set (an hour or two), these plates are innoculated in the same way as Method 1. These plates yield from . 5 to 1 gm of nematodes per plate after 3 to 4 days at room temperature. This method will provide quantities of worms for biochemical purposes at approximately 1/10 to 1/100 the cost per gram of Method 1.