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Main sol. 1011

Internal Solution-ID: S2058

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.674 507.762
K2HPO4 0.14 g 0.138 0.795
CaCl2 x 2 H2O 0.14 g 0.138 0.942
MgSO4 x 7 H2O 3.40 g 3.363 13.644
MgCl2 x 6 H2O 4.18 g 4.135 20.337
KCl 0.33 g 0.326 4.378
NH4Cl 0.25 g 0.247 4.623
Fe(NH4)2(SO4)2 x 6 H2O 0.01 g 0.01 -
Modified Wolin's mineral solution 10.00 ml - -
NaHCO3 1.50 g 1.484 17.661
Na2S2O3 x 5 H2O 1.50 g 1.484 5.978
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, thiosulfate and vitamins), then sparge medium with 80% N2 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 bicarbonate, thiosulfate and vitamins to the autoclaved medium from sterile anoxic stock solutions. Solutions of vitamins and thiosulfate are sterilized by filtration and stored under N2, whereas the solution of bicarbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust pH of complete medium to 6.7.
2 After inoculation pressurize vessels to 0.5 bar overpressure with sterile 80% N2 and 20% CO2 gas mixture and add sterile air in an amount that is equivalent to a volume of 20% of the headspace.
1011 SULFURIMONAS MJ MEDIUM