EMM−N with 1% Glucose

Reagent	Quantity (for 1 L)	Final concentration
Potassium hydrogen phthalate	3 g	14.7 mm
Na₂HPO₄	2.2 g	15.5 mm
Glucose	10 g	1%
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 EMM−N with 1% glucose 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 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).