Equipment needed: Filter
Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Baker's yeast | 2.50 | g | 2.5 | - | |
| CaCl2 x 2 H2O | 1.36 | g | 1.36 | 9.251 | |
| Vitamin B12 | 0.50 | mg | 5.0e-4 | 3.7e-4 | |
|
Agar
(Difco) |
8.00 | g | 8 | 23.786 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Sterilize vitamin B12 separately by filtration. Prepare and store yeast cells as autoclaved stock suspension (5 g baker's yeast/100 ml distilled water, adjust pH to 6.5 and autoclave). | |||||
| 2 Adjust pH of medium to 7.2 with KOH before as well as after autoclaving and cooling to 50°C (use pH-indicator paper). | |||||
| 3 For suspension of freeze-dried cells from ampoules add about 0.5 - 1.0 ml medium MD1 (per liter: casiton, 3.0 g; CaCl2 x 2 H2O, 0.7 g; MgSO4 x 7 H2O, 2.0 g) to the vial with freeze dried material. | |||||
Media with this solution
| 9a | VY/2, REDUCED MEDIUM |