Main sol. 829

Equipment needed: Hungate tubes Filter Autoclave Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Sea Salt (SIGMA, must not show signs of caking)	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
Sulfur , powdered	10.00	g	9.95	310.267
Na₂CO₃	0.50	g	0.498	4.694
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 sulfur, carbonate, 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 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 growth-stimulating factors from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate and Na-dithionite from sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Vitamins and Na-dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.0, if necessary.
2 After inoculation use 1 bar overpressure of sterile 80% H₂ and 20% CO₂ gas mixture.

829	DESULFUROBACTERIUM MEDIUM