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Main sol. 829b

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Internal Solution-ID: S1682

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA) 		30.00 g 29.851 -
NH4Cl 1.00 g 0.995 18.602
KH2PO4 0.35 g 0.348 2.559
MES [2-(N-morpholino) ethane sulfonic acid] 1.95 g 1.94 9.938
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 0.50 g 0.498 4.694
Yeast extract 0.50 g 0.498 -
Seven vitamins solution 1.00 ml - -
Growth-stimulating factors 1.00 ml - -
Na-dithionite solution (5% w/v) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, yeast extract, vitamins, growth-stimulating factors and Na-dithionite), boil medium for 1 min, then cool to room temperature under 80% H2 and 20% CO2 gas atmosphere. Adjust pH to 6.0 and dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add yeast extract, vitamins, and growth-stimulating factors from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate and Na-dithionite from sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins and Na-dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.5, if necessary.
2 After inoculation use 2 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.
829b THERMODESULFOBACTERIUM HYDROGENIPHILUM MEDIUM