1. Suspend 41 grams of the medium in 900 mL of purified water.
2. Heat with frequent agitation and boil for one minute to completely dissolve the medium.
3. Autoclave at 121°C for 15 minutes. Cool to 45 – 50°C.
4. Add 100 mL of NCM4017* Egg Yolk Emulsion, 50%, and the desired antimicrobic supplement: TSC Agar: Add 2 vials of NCM4022-0.5* TSC Perfringens Supplement, each reconstituted using 5 mL sterile deionized/RO water.

Overlay Agar

1. Suspend 41 grams of the medium in one liter of purified water.
2. Heat with frequent agitation and boil for one minute to completely dissolve the medium.
3. Autoclave at 121°C for 15 minutes. Cool to 45 – 50°C.
4. Overlay agar does not get supplemented with Egg Yolk Emulsion. Overlay agar is held at 45 -50°C until used and should be poured right after inoculation.

*Larger vials may be available. Please see appropriate supplement data sheet for availability and preparation instructions.