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Main sol. 1020

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Internal Solution-ID: S2072

Equipment needed: Autoclave Hungate tubes Filter Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaNO3 0.2 g 0.2 2.348
KH2PO4 0.2 g 0.2 1.467
NH4Cl 0.2 g 0.2 3.732
MgCl2 x 6 H2O 0.4 g 0.399 1.964
KCl 0.2 g 0.2 2.677
CaCl2 x 2 H2O 0.1 g 0.1 0.679
Trace element solution SL-4 2.0 ml - -
Na2S2O3 x 5 H2O 2.5 g 2.495 10.053
Distilled water 1000.0 ml - -
1 Dissolve ingredients, except sodium thiosulfate, then sparge solution with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 7.0, then distribute medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Prior to inoculation, add thiosulfate from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
2 After inoculation, pressurize culture vessels to 0.5 bar overpressure with sterile 100% H2 gas.
1020 SULFURICURVUM (MBM) MEDIUM