|Trypticase (tryptic) soy agar||40 g|
|Yeast extract||5 g|
|L-Cysteine (dissolved in 5 ml 1 N NaOH)||0.4 g|
|Distilled water||l liter|
Heat with agitation to dissolve agar. Adjust pH to 7.5 ± 0.2. Autoclave 15 min at 121°C. Cool to 50°C.
Suspend 1 g hemin in 100 ml distilled water. Autoclave 15 min at 121°C. Refrigerate at 4°C.
Vitamin K1 solution
Dissolve 1 g vitamin K1 (Sigma Chemical Co., St. Louis, MO) in 100 ml 95% ethanol. Solution may require 2-3 days with intermittent shaking to dissolve. Refrigerate at 4°C.
To 1 liter base add 0.5 ml hemin solution and 1 ml Vitamin K1 solution. Mix and pour 20 ml portions into 15 × 100 mm petri dishes. Medium must be reduced before inoculation by 24 h anaerobic incubation in anaerobic glove box or GasPak jar.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.