Nitrogen-Rich Minimal Sporulation Liquid (MSL+N) Medium

Recipe

Reagent	Quantity (for 1 L)	Final concentration
Glucose	10 g	55.5 mm
(NH₄)₂SO₄	2 g	15.12 mm
KH₂PO₄	1 g	7.3 mm
NaCl	0.1 g	1.7 mm
MgSO₄ · 7H₂O	0.2 g	0.81 mm
CaCl₂ · 6H₂O (0.1 m)	1 mL	0.1 mm
Minerals (10,000×)	0.1 mL	1×
Vitamins (1000×)	1 mL	1×
Adenine (optional; see below)	0.225 g	1.7 mm
Leucine (optional; see below)	0.225 g	1.7 mm
Uracil (optional; see below)	0.225 g	2.0 mm
H₂O	to 1 L
To allow the growth of auxotrophic strains, include leucine, adenine, and uracil. (Additional amino acids should be included if the strain used carries a distinct auxotrophy [e.g., his3Δ].) Filter-sterilize using a 0.22-µm pore size membrane. (For live cell imaging, filter-sterilized medium has lower background fluorescence than autoclaved medium.) Store at 4°C.