How long do NGM plates last?
To seed plates, dispense 100-120 μl of the OP50 onto the 60 mm diameter NGM Petri plate and swirl to spread it. Incubate these seeded plates overnight at room temperature for the lawn to grow. These seeded plates can now be stored at 4° C for up to 3 weeks.
What is NGM Lite?
NGM-Lite All components mixed with 1L water and then autoclaved: 2.0g NaCl, 4.0g Bactotryptone, 3.0 g KH2PO4, 0.5g K2HPO4, 8mg Cholesterol, 20g Agar. You will probably see some particles after autoclaving, but their density is pretty low and they don’t interfere with optics on the dissecting scope.
What is NGM plate?
elegans is maintained in the laboratory on Nematode Growth Medium (NGM) agar which has been aseptically poured into petri plates. A constant amount of agar in the plates reduces the need for refocusing the microscope when you switch from one plate to another.
What are seeded NGM plates used for?
Seeding Plates to use for Culturing Worms You will need the following: NGM plates (determine the number you need to seed, then add a few extra) Bacterial culture in LB broth (made as above) 5 ml or 10 ml sterile disposable pipet(s)
How do you make NGM agar plates?
Nematode Growth Medium (NGM)
- For solid NGM, mix 3 g of NaCl, 2.5 g of peptone, and 20 g of agar and bring to 1 L with H2O.
- Autoclave for 1 h.
- Let cool for 1 h in a 55°C water bath.
What is in NGM agar?
For the cultivation of nematodes like C. elegans. Contains: 0.25% Tryptone (2.5 g/L), 0.3% Sodium Chloride (NaCl) (3.0 g/L), 1.5% Agar, 1 mM Calcium Chloride, 1 mM Magnesium Sulfate, 25 mM Potassium Phosphate (KPO4) pH 6.0, and 5 ®µg/mL Cholesterol.
How do you make NGM?
How long does E coli OP50 need to grow before adding worms to the plate?
coli OP50 lawn to grow overnight at room temperature or at 37°C for 8 hours (cool plates to room temperature before adding worms). Seeded plates stored in an air-tight container will remain usable for 2-3 weeks.
How do I keep C. elegans alive?
elegans stocks. Caenorhabditis elegans can be frozen and stored indefinitely in liquid nitrogen (−196 °C) (Brenner, 1974). The keys to a successful freeze are using animals at the correct stage of development, the addition of glycerol to the freezing media, and a gradual cooling to -80°C.