| l-Serine
|
2.1 g |
20 mm |
| Potassium hydrogen phthalate |
3 g |
14.7 mm |
| Na2HPO4 |
2.2 g |
15.5 mm |
| Glucose |
20 g |
111 mm |
| Salt stock (50×)
|
20 mL |
1× |
| Vitamins (1000×)
|
1 mL |
1× |
| Minerals (10,000×)
|
0.1 mL |
1× |
| H2O
|
to 1 L |
|
| Prepare 1 L of EMMSer by combining the reagents above. To prepare solid medium, include agar (20 g/L). 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).
|
