When growing C. elegans, large amounts can be grown in different mediums. The most popular is liquid medium. C. elegans are usually grown on S Medium using concentrated doses of E. coli bacteria as their food. Starting off, it is wise to grow just one generation of worms before harvesting them. After the worms produce multiple generations, harvesting should be done.
One should make a concentrated plate of bacteria. This plate can be store around 4/5°C for several weeks. One can store the plates in a subzero freezer (-70°C) indefinitely. Generally speaking, it is a sound idea to have ample amounts of E. coli available for the C.elegans. However, the amount of food the worms will need will depend on the starting inoculate of worms and the length of time the worms are grown.
Here are some of the common reagents used in growing C.elegans
- NaCl, K2 HPO4, 6 g KH2PO4, 1 ml cholesterol , water. Sterilize it by using an autoclave machine.
- 1 M Potassium citrate pH 6.0
- Some trace metals solutions
- 1 M CaCl2
- 2 or 3 large plates of C. elegans
- food, ie. concentrated E. coli
Inoculate the Medium with a does of E. coli from an previously made culture. Make sure you wash each plates of C. elegans properly. Shake the flask using a shaker at 20°C for a predetermined amount of time. Make sure the flask is shaken vigorously to ensure the culture is well oxygenated. Monitor each culture by examining them under a microscope. When examining the C.elegans, if their food supply is depleted, it is time to replenish the supply. However one must also harvest them, which is usually done on the 5th day. Sterilize the harvesting tool, and transfer 5 or so C.elegans onto a new plate