Nematode growth media (NGM) plates

K₂HPO₄ (1 M stock solution)

KH₂PO₄ (1 M stock solution)

NaCl

Bacto peptone

Agar

Agarose

MgSO₄ (1 M stock solution)

CaCl₂ (1 M stock solution)

Cholesterol stock solution (5 mg/mL in ethanol)

• 1. Prepare potassium phosphate buffer for use in Step 5 by mixing 132 mL of K₂HPO₄ (1 M) with 868 mL of KH₂PO₄ (1 M). Filter-sterilize.

• 2. Dissolve 3 g/L NaCl and 2.5 g/L bacto peptone in H₂O.

• 3. Add 17 g/L agar (for 6- and 10-cm plates) or 10 g/L agar and 10 g/L agarose (for 14.5-cm plates).

• 4. Autoclave; then cool the solution to 50°C.

• 5. Add the following components:

  • 25 mL potassium phosphate buffer (prepared in Step 1)

  • 1 mL MgSO₄ (1 M)

  • 1 mL CaCl₂ (1 M)

  • 1 mL cholesterol (5 mg/mL in ethanol)

• 6. Mix the solution well and pour into Petri dishes.