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BAM Media M11: Anaerobe Agar

January 2001

Bacteriological Analytical Manual

Anaerobe Agar

Trypticase (tryptic) soy agar40 g
Agar5 g
Yeast extract5 g
L-Cysteine (dissolved in 5 ml 1 N NaOH)0.4 g
Distilled waterl liter


Heat with agitation to dissolve agar. Adjust pH to 7.5 ± 0.2. Autoclave 15 min at 121°C. Cool to 50°C.



Hemin solution

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.


Final medium

To 1 liter base add 0.5 ml hemin solution and 1 ml Vitamin K1 solution. Mix and pour 20 ml portions into 15 x 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.