# 1513: SULFURIMONAS GOTLANDICA MEDIUM ## Main sol. 1513 5.55 g NaCl 2.29 g MgCl2 x 6 H2O 0.34 g CaCl2 x 2 H2O 0.15 g KCl 0.91 g Na2SO4 1.01 g KNO3 2.38 g HEPES buffer (SIGMA) 0.05 g NH4Cl 8 ml SrCl2 (0.1% w/v) 6 ml H3BO3 (0.1% w/v) 4 ml NaF (0.1% w/v) 1 ml K2HPO4 (0.1% w/v) 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 0.2 g NaHCO3 2.48 g Na2S2O3 x 5 H2O 1 ml **Wolin's vitamin solution (10x)** 980 ml Distilled water 1. Dissolve ingredients (except bicarbonate, thiosulfate and vitamins) and adjust pH to 7.3. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes) and autoclave. Add thiosulfate and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of vitamins and thiosulftae should be sterilized by filtration. Adjust pH of the complete medium to 7.3 - 7.5, if necessary. 2. After inoculation pressurize vials to 0.7 bar overpressure with sterile 100% N2 gas. ## 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