# 981: GEOTHERMOBACTER MEDIUM ## Main sol. 981 19 g NaCl 9 g MgCl2 x 6 H2O 0.15 g MgSO4 x 7 H2O 0.3 g CaCl2 x 2 H2O 0.5 g KCl 0.42 g KH2PO4 0.1 g (NH4)2SO4 0.05 g NaBr 0.02 g SrCl2 x 6 H2O 1.01 g KNO3 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 1 g Na2CO3 0.1 g Yeast extract 1.34 g DL-Malic acid 1 ml **Wolin's vitamin solution (10x)** 0.05 g L-Cysteine HCl x H2O 1000 ml Distilled water 1. Dissolve ingredients (except carbonate, yeast extract, malic acid, vitamins and cysteine) and sparge medium with 80% N2 and 20% CO2 gas mixture 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 yeast extract, malic acid, vitamins (sterilized by filtration), and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The pH of the complete medium should be 6.2. ## 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. ## Selenite-tungstate solution 0.5 g NaOH 3 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 x 2 H2O 1000 ml Distilled water ## 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