Low-Glucose (0.5%) EMM2

Recipe

Reagent	Quantity (for 1 L)	Final concentration
Ammonium chloride	5 g	93.5 mm
Potassium hydrogen phthalate	3 g	14.7 mm
Na₂HPO₄	2.2 g	15.5 mm
Glucose	5 g	28 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 low-glucose (0.5%) EMM2 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). If desired, add 12 μm thiamine (364 μL of a filter-sterilized stock solution of 10 mg/mL in H₂O) to repress expression from nmt1-derived promoters (Maundrell 1990). (For liquid medium, low pressure and a short cycle is essential to avoid caramelization of glucose and breakdown of vitamins and minerals. Store at 4°C.