| Glucose |
10 g |
55.5 mm |
| Arginine hydrochloride |
2 g |
9.5 mm |
| KH2PO4 |
1 g |
7.3 mm |
| NaCl |
0.1 g |
1.7 mm |
| MgSO4 · 7H2O
|
0.2 g |
0.81 mm |
| CaCl2 · 6H2O
|
0.15 g |
0.68 mm |
| Minerals (10,000×)
|
0.1 mL |
1× |
| Vitamins (1000×)
|
2 mL |
1× |
| H2O
|
to 1 L |
|
| Prepare minimal sporulation liquid (MSL) medium (Egel et al. 1994) by combining the reagents in the list above. To prepare solid minimal sporulating agar (MSA) medium, include 20 g/L of agar.
Sterilize by autoclaving at 10 psi for 10 min (for liquid medium) or 15 min (for solid medium). (For liquid medium, low pressure
and a short cycle is essential to avoid caramelization of glucose and breakdown of vitamins and minerals. For live cell imaging,
filter-sterilized medium has lower background fluorescence than autoclaved medium. Use a 0.22-µm pore size for filtration.)
Store at 4°C. If desired, add 12 µm thiamine (364 µL of a filter-sterilized stock solution of 10 mg/mL on H2O) after autoclaving to fully repress expression from nmt1-derived promoters (Maundrell 1990).
|
