# J1204: SPOROMUSA ACIDOVORANS MEDIUM ## Main sol. J1204 0.5 g NH4Cl 0.5 g MgSO4 x 7 H2O 0.25 g CaCl2 x 2 H2O 2.25 g NaCl 1 ml **FeCl2 solution** 1 ml **Trace element solution** 1 ml **Se/W solution** 2 ml FeSO4 x 7 H2O (in 0.1 N H2SO4, 0.1%) 2 g Yeast extract 2 g Casitone (BD-Difco) 0.5 mg Resazurin 830 ml Distilled water 1. Solution B: 0.35 g K2HPO4 0.23 g KH2PO4 100 ml Distilled water 2. After autoclaving, combine solutions A and B, then add the following solutions (filter-sterilized): 10 ml **Trace vitamins** 10 ml D-Fructose (w/v, 40%) 50 ml NaHCO3 (8%) 3. To complete the medium, aseptically and anaerobically distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a N2-CO2 (4:1, v/v) gas stream and seal with butyl rubber stoppers. Prior to use, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere): 6 ml L-Cysteine HCl x H2O (5%) 6 ml Na2S x 9 H2O (5%) 4. Solution A: 5. Autoclave under a N2 gas atmosphere. 6. Autoclave under a N2 gas atmosphere. 7. Adjust pH to 6.5-7.0, if necessary. ## FeCl2 solution 10 ml HCl (7.7 M, 25%) 1.5 g FeCl2 x 4 H2O 990 ml Distilled water ## Trace element solution 70 mg ZnCl2 100 mg MnCl2 x 4 H2O 6 mg H3BO3 190 mg CoCl2 x 6 H2O 2 mg CuCl2 x 2 H2O 24 mg NiCl2 x 6 H2O 36 mg Na2MoO4 x 2 H2O 1000 ml Distilled water ## Se/W solution 4 mg Na2SeO3 4 mg Na2WO4 x 2 H2O 1000 ml Distilled water ## Trace vitamins 2 mg Biotin 2 mg Folic acid 10 mg Pyridoxine hydrochloride 5 mg Thiamine HCl 5 mg Riboflavin 5 mg Nicotinic acid 5 mg Calcium pantothenate 0.1 mg Vitamin B12 5 mg p-Aminobenzoic acid 5 mg Lipoic acid 1000 ml Distilled water