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.