Nitrogen-Deprived Minimal Sporulation Liquid (MSL−N) Medium with Agarose
Combine 10 mL of nitrogen-deprived minimal sporulation liquid (MSL–N) medium with 0.2 g of agarose. Place in a microwave oven at high power until the agarose dissolves (∼2 min). Depending on the nature of the experiment, add P- and/or M-factor pheromones to the melted agarose solution to the desired final concentration (e.g., 0.001–1.0 µg/mL). (The pheromones are typically prepared as 1 mg/mL stock solutions in methanol, so use pheromone-free methanol for control pads.) Store 0.5-mL aliquots in microcentrifuge tubes at room temperature.
