Casitone 10.00 g Yeast extract 2.50 g Glucose 5.00 g MgSO4 x 7 H2O 45.00 mg CaCl2 x 2 H2O 90.00 mg K2HPO4 0.45 g KH2PO4 0.45 g NaCl 0.9 g Resazurin 1.0 mg Distilled water 1000.00 ml NaHCO3 4.00 g L-Cysteine-HCl 1.00 g Hemin 10.00 mg Volatile fatty acids: Acetic acid 1.90 ml Propionic acid 0.70 ml iso-Butyric acid 90.00 μl n-Valeric acid 100.00 μl iso-Valeric acid 100.00 μl Vitamin solution: Biotin 2.00 mg Folic acid 2.00 mg Pyridoxine-HCl 10.00 mg Thiamine-HCl x 2 H2O 5.00 mg Riboflavin 5.00 mg Nicotinic acid 5.00 mg D-Ca-pantothenate 5.00 mg Vitamin B12 0.10 mg p-Aminobenzoic acid 5.00 mg Lipoic acid 5.00 mg Distilled water 1000.00 ml Dissolve ingredients except NaHCO3, hemin, cysteine in water and boil for 10 min, then cool to room temperature while gassing with 100% CO2.Add the NaHCO3, hemin, cysteine and adjust pH to 6.7 -6.8. Distribute under N2 into Hungate tubes (10 ml) and autoclave. Filter sterilize vitamin solution and aseptically add 100 μl per Hungate tube.
