# 1255: THERMOVENABULUM MEDIUM ## Main sol. 1255 1 g NaCl 1 g NH4Cl 0.1 g MgCl2 x 6 H2O 0.1 g CaCl2 x 2 H2O 0.38 g KH2PO4 0.75 g K2HPO4 10 ml **Trace element solution SL-10** 2 g Yeast extract (OXOID) 2 g Tryptone (BD Bacto) 0.5 ml FeSO4 (0.1% w/v in 0.1 N H2SO4) 3.8 g PIPES (SIGMA) 0.5 ml Sodium resazurin (0.1% w/v) 2 g D-Glucose 1 ml **Wolin's vitamin solution (10x)** 0.5 g Na2S x 9 H2O 1000 ml Distilled water 1. Dissolve ingredients (except glucose, vitamins and sulfide), adjust pH to 7.0 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add glucose, vitamins and sulfide from sterile anoxic stock solution prepared under 100% N2 gas. Vitamins are sterilized by filtration. The pH of the complete medium should be 7.0. ## Trace element solution SL-10 10 ml HCl (25%) 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. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. ## Wolin's vitamin solution (10x) 20 mg Biotin 20 mg Folic acid 100 mg Pyridoxine hydrochloride 50 mg Thiamine HCl 50 mg Riboflavin 50 mg Nicotinic acid 50 mg Calcium D-(+)-pantothenate 1 mg Vitamin B12 50 mg p-Aminobenzoic acid 50 mg (DL)-alpha-Lipoic acid 1000 ml Distilled water