B-11680

KCl 0.33 g MgSO4 x 7 H2O 3.45 g MgCl2 x 6 H2O 2.75 g NaCl 20.00 g CaCl2 x 2 H2O 0.15 g NH4Cl 0.25 g K2HPO4 0.15 g Fe(NH4)2(SO4)2 x 6 H2O 0.01 g Yeast extract 0.10 g Trace elements solution SL6 (see medium 27) 1.00 ml Na-resazurin solution (0.1% w/v) 0.50 ml Na2CO3 1.00 g D-Glucose 2.00 g Vitamins solution (see medium 141) 10.00 ml L-Cysteine-HCl x H2O 0.30 g Na2S x 9 H2O 0.30 g Distilled water 1000.00 ml Dissolve ingredients except carbonate, glucose, vitamins, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, glucose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 6.0 - 6.5, if necessary.