KH2PO4 0.30 g K2HPO4 0.30 g NH4Cl 1.00 g NaCl 1.00 g KCl 0.10 g MgCl2 x 6 H2O 0.50 g CaCl2 x 2 H2O 0.10 g Trace element solution SL-10 1.00 ml Yeast extract (OXOID) 1.00 g Sodium resazurin (0.1% w/v) 0.50 ml Sulfur (powdered) 10.00 g L-Cysteine HCl x H2O 0.50 g Trypticase peptone (BD BBL) 2.00 g Na2-fumarate 3.20 g Na2CO3 1.00 g Na2S x 9 H2O 0.50 g Distilled water 1000.00 ml Dissolve ingredients (except sulfur, cysteine, peptone, fumarate, carbonate and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissove cysteine, then dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and autoclave at 121°C for 20 min. Add peptone, fumarate and sulfide 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. Adjust pH of the complete medium to 7.0. For DSM 24444: Omit Trypticase peptone and fumarate. Increase amount of yeast extract to 2.0 g/l and supplement medium with 0.2 g/l Na-acetate and 0.6 g/l DL-lactate added after autoclaving from sterile anoxic stock solutions prepared under 100% N2 gas. For DSM 25546: Omit Trypticase peptone and fumarate. Increase amount of yeast extract to 2.0 g/l, supplement medium with 0.2 g/l Na-acetate and add 3.0 g/l D-xylose after autoclaving from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration. For DSM 29926: Supplement medium with 6.0 g/l D-glucose added to the medium after autoclaving from a sterile anoxic stock solution prepared under 100% N2 gas. Trace element solution SL-10 (from medium 320) HCl (25%) 10.00 ml FeCl2 x 4 H2O 1.50 g ZnCl2 70.00 mg MnCl2 x 4 H2O 100.00 mg H3BO3 6.00 mg CoCl2 x 6 H2O 190.00 mg CuCl2 x 2 H2O 2.00 mg NiCl2 x 6 H2O 24.00 mg Na2MoO4 x 2 H2O 36.00 mg Distilled water 990.00 ml First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
