Main sol. 829a

Equipment needed: Filter Autoclave Hungate tubes Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	30.00	g	29.412	503.281
MgCl₂ x 6 H₂O	3.00	g	2.941	14.467
CaCl₂ x 2 H₂O	0.15	g	0.147	1
KCl	0.50	g	0.49	6.575
NH₄Cl	0.50	g	0.49	9.164
MES [2-(N-morpholino) ethane sulfonic acid]	1.95	g	1.912	9.792
KH₂PO₄	0.20	g	0.196	1.441
Sodium resazurin (0.1% w/v)	0.50	ml	4.9e-4	0.002
Na₂S₂O₃ x 5 H₂O	2.50	g	2.451	9.876
Neutralized sulfide solution 3% (w/v)	20.00	ml	-	-
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except thiosulfate and sodium sulfide), adjust pH to 6.0, boil medium for 1 min, then cool to room temperature under 80% H₂ and 20% CO₂ gas atmosphere. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add sodium thiosulfate from an anoxic stock solution prepared under 100% N₂ gas and sterilized by filtration. Reduce medium with a sterile, neutralized solution of sodium sulfide prepared under 100% N₂ gas . Adjust pH of complete medium to 6.0, if necessary.
2 After inoculation pressurize cultivation vessels to 2 bar overpressure using sterile 80% H₂ and 20% CO₂ gas mixture.

829a	DESULFUROBACTERIUM ATLANTICUM MEDIUM