Per Liter of medium (~75 plates):
- 975 mL Water
- 3 g NaCl
- 2.5 g Peptone (Fisher BP1420-500 $78.80)
- 17 g Bactoagar
This is what I did by mistake: per 1.5 L (112 plates):
- 1462 mL Water
- 18 g NaCl
- 15 g Peptone (Fisher BP1420-500 $78.80)
- 25.5 g Bactoagar (haven't added it yet)
So I labeled it NGM 4X.
I made 4 liters by diluting 975 mL 4xNGM with 975 x 3 mL water, then added 68 g bactoagar.
Stir flask to distribute ingredients.
Autoclave with stir bar inside
Cool to 55C in a 55C water bath
Add per L (see recipes in stock solutions):
- 1 mL cholesterol (5 mg/mL in 95% EtOH)
- 1 mL CaCl2 (1 M, STERILE)
- 1 mL MgSO4 (1 M, STERILE)
- 25 mL K-phosphate buffer (1M, pH 6.0, STERILE1)
Swirl flask to mix
Dispense 10 mL into each 60mm dish.
Stack 10 high
Let stand for ~48 hours for condensation to evaporate
Pack in sterilized plastic boxes.
3.3 mL K2HPO4 + 21.7 mL KH2PO4 ↩︎