Medium 33: BRAIN HEART INFUSION AGAR (SUPPLEMENTED)
Brain Heart Infusion (Oxoid) 37 g
Yeast extract 5 g
Resazurin solution (0.025%) 4 ml
Agar 25 g
Distilled water 1000 ml
pH 7.0
Sterilize at 121°C for 15 min.
Boil, cool, add sterile:
L-Cysteine.HCl 0.5 g
Hemin solution* 10 ml
Vitamin K1** 0.2 ml
*Hemin solution: Dissolve 50 mg hemin in 1ml 1N
NaOH: make 100 ml with distilled water. Sterilize
at 121°C for 15 min.
**Vitamin K1 solution: Dissolve 0.15 ml of vitamin
K1 in 30 ml of 95% ethanol. Do not sterilize
since it is added to media before autoclaving.
Keep under refrigeration. Store in brown bottle.
Discard after 1 month.