# J1009: CLOSTRIDIUM SWELLFUNIANUM MEDIUM ## Main sol. J1009 0.33 g NH4Cl 0.33 g KCl 0.33 g KH2PO4 0.5 g Yeast extract (BD-Difco) 3 g Tryptone (BD-Difco) 1 ml **Trace element solution SL-10** 10 ml **Trace vitamins** 33 ml CaCl2 x 2 H2O (1%) 16.5 ml MgCl2 x 6 H2O (2%) 0.5 g L-Cysteine HCl x H2O 1 ml Resazurin (1%) 1000 ml Distilled water 1. Prepare the medium anaerobically under a N2-CO2 (4:1, v/v) gas mixture. Distribute the medium into culture vessels (e.g., 5.0 ml in Hungate tubes) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After autoclaving, add per 5.0 ml the following solutions from anaerobic stocks (autoclaved or *filter-sterilized and stored under a N2 atmosphere): 0.05 ml Na2S x 9 H2O (w/v, 3%) 0.05 ml NaHCO3 (w/v, *, 10%) 0.1 ml Glucose (w/v, 20%) 2. Adjust pH to 7.3. ## Trace element solution SL-10 10 ml HCl (25%, 7.7 M) 1.5 g FeCl2 x 4 H2O 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 990 ml Distilled water 1. Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. ## 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