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Main sol. 1011c

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Internal Solution-ID: S2061

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.644 507.26
K2HPO4 0.14 g 0.138 0.794
CaCl2 x 2 H2O 0.14 g 0.138 0.941
MgSO4 x 7 H2O 3.40 g 3.36 13.631
MgCl2 x 6 H2O 4.18 g 4.13 20.317
KCl 0.33 g 0.326 4.374
NH4Cl 0.25 g 0.247 4.618
Fe(NH4)2(SO4)2 x 6 H2O 0.01 g 0.01 -
Modified Wolin's mineral solution 10.00 ml - -
Na2CO3 1.50 g 1.482 13.985
Na-pyruvate 0.50 g 0.494 4.49
Na-lactate 0.50 g 0.494 4.409
Yeast extract 0.10 g 0.099 -
Na2S2O3 x 5 H2O 1.50 g 1.482 5.972
Wolin's vitamin solution (10x) 1.00 ml - -
Na-dithionite solution (5% w/v) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, pyruvate, lactate, yeast extract, thiosulfate, vitamins and dithionite), then sparge medium with 80% H2 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 up to a volume of 20% and autoclave. Add pyruvate, lactate, yeast extract, thiosulfate and vitamins to the autoclaved medium from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate and dithionite from sterile anoxic stock solutions prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins, thiosulfate and dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.7.
2 After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
1011c DESULFOTHERMUS MJ MEDIUM (H2/CO2)