05 - Heavy Metal: Effects of Environmental Toxins on C. elegans
Module I: Preparation of C. elegans Food Source (“Seeding” the Plates) Excerpts from Edvo-Kit #856 In this module, you will seed two petri plates with a favorite C. elegans’ food ( E. coli OP50 strain bacteria).
SPREAD culture
NGM Plates
100 µL OP50
SPREAD culture
COVER & INCUBATE 24 hours.
00 µL
OP50
ROOM TEMP.
1. OBTAIN two Nematode Growth Medium (NGM) plates, the OP50 culture, a small transfer pipet, and a sterile inoculat- ing loop from your instructor. LABEL the bottom of both plates with your group number or names.
HINTS for Step 3: • Avoid gouging or scratching the agar
surface as this can affect visibility as well as worm movement. • Widely spreading the bacteria creates a larger lawn for the worms but stop just before the plate's edge. This discour- ages the worms from crawling up the plate's sides and drying out.
2. While maintaining sterile technique, ADD two drops (100 µL) of OP50 culture to each plate.
3. Using the loop, SPREAD the culture over the entire surface of the NGM plates. COVER . 4. INCUBATE the plates inverted and at room temperature for 24 hours. NOTE: Seeded OP50 plates can also be prepared overnight (~12 hours) by incubating at 37°C.
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