| Bacto Heart Infusion Broth (Difco) | 17.5 | g | | Horse serum* | 200 | ml | | 15% Yeast Extract Solution (Gibco 0689) | 150 | ml | | Sucrose | 40 | g | | Phenol red | 40 | mg | | Distilled water | 650 | ml | | | | | | | | | | *Heat-inactivated at 56℃ for 30 min. | | | Dissolve 17.5 g of heart infusion broth and 40 mg of phenol red in 450 ml of water (sterilized by autoclaving at 121℃ for 15 min). Dissolve sucrose in 200 ml of water (sterilized by filtration). Aseptically add the horse serum, yeast extract solution, sterile sucrose, and 1 ml of Penicillin G (200,000 U/ml, sterilized by filtration) to the cooled heart infusion broth. Final pH 7.4 +/- 0.2. |
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