NH4Cl 0.50 g KH2PO4 0.20 g MgCl2 x 6 H2O 0.10 g KCl 0.20 g Trace element solution SL-10 1.00 ml Selenite-tungstate solution 1.00 ml Na2CO3 1.00 g NaHCO3 7.60 g NaCl 10.00 g Yeast extract 0.20 g Cellobiose 3.00 g Na2S x 9 H2O 0.50 g Distilled water 1000.00 ml Dissolve ingredients except cellobiose and sulfide, then 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. Add cellobiose after autoclaving from an anoxic stock solution prepared under 100% N2 gas sterilized by filtration and sulfide from a sterile anoxic stock solution autoclaved under 100% N2 gas. Adjust pH of the complete medium to pH 8.8-9.0. 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. Selenite-tungstate solution (from medium 385) NaOH 0.50 g Na2SeO3 x 5 H2O 3.00 mg Na2WO4 x 2 H2O 4.00 mg Distilled water 1000.00 ml
