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Main sol. 829d

Internal Solution-ID: S1684

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA)
30.00 g 29.412 -
NH4Cl 0.30 g 0.294 5.499
KH2PO4 0.35 g 0.343 2.521
MES [2-(N-morpholino) ethane sulfonic acid] 1.95 g 1.912 9.792
Na-acetate 0.16 g 0.157 1.912
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Sulfur , powdered 10.00 g 9.804 305.704
Na2CO3 0.50 g 0.49 4.625
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.49 2.041
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, carbonate, vitamins and sulfide), 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 under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. Autoclave at a temperature of 110°C for 20 min! Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins are sterilized by filtration. Adjust pH of the complete medium to 6.0 - 6.5, if necessary.
2 After inoculation add 1 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.
829d DESULFUROBACTERIUM INDICUM MEDIUM