Table 1:
Recipes for NGM plates, M9 buffer, and bleach solution.
| M9 buffer (1 L) | |||
|---|---|---|---|
| dH2O | 1,000 mL | ||
| NaCl | 5 g | ||
| KH2PO4 | 3 g | ||
| Na2HPO4 | 6 g | ||
| 1 M MgSO4 | 1 mL *add after autoclaving | ||
| Split between 2 bottles: 500 mL each. Autoclave on liquid cycle (15 min exposure) | |||
| Bleach solution (50 mL) | |||
| dH2O | 36 mL | ||
| Bleach | 14 mL | ||
| 10 N NaOH | 800 μL | ||
| Standard Nematode Growth Media (NGM) plates | |||
| 1 L | 0.5 L | 0.25 L | |
| NaCl | 3 g | 1.5 g | 0.75 g |
| Bacto-agar | 17 g | 8.5 g | 4.25 g |
| Bacto-peptone | 2.5 g | 1.25 g | 0.625 g |
| a. Autoclave on liquid cycle (45 min exposure), allow to cool to ~60 °C and then use sterile technique and add the following: | |||
| 1 L | 0.5 L | 0.25 L | |
| 1 M CaCl2 | 1 mL | 0.5 mL | 0.25 mL |
| 1 M MgSO4 | 1 mL | 0.5 mL | 0.25 mL |
| 1 M KPO4 | 25 mL | 12.5 mL | 6.25 mL |
| 5 mg/mL cholesterol | 1 mL | 0.5 mL | 0.25 mL |
| b. Swirl to mix thoroughly after each addition. After all additions are made, pour plates | |||