Sporulation Agar (SPA) Medium

Reagent	Quantity (for 1 L)	Final concentration
Glucose	10 g	55.5 mm
KH₂PO₄	1 g	7.3 mm
Vitamins (1000×)	1 mL	1×
Agar	20 g	20 g/L
H₂O	to 1 L
To prepare SPA medium (Munz and Leupold 1970), combine the ingredients listed above. Sterilize by autoclaving at 10 psi for 15 min. 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).

REFERENCES

1. Maundrell K
Maundrell K. 1990. Nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine. J Biol Chem 265: 10857–10864.
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1. Munz P,
2. Leupold U
Munz P, Leupold U. 1970. Characterization of ICR-170-induced mutations in Schizosaccharomyces pombe. Mutat Res 9: 199–212.
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