Main sol. 977

Equipment needed: Anaerobic workbench Hungate tubes Filter Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NH₄Cl	0.33	g	0.33	6.169
MgCl₂ x 6 H₂O	0.33	g	0.33	1.623
CaCl₂ x 2 H₂O	0.33	g	0.33	2.245
KCl	0.33	g	0.33	4.427
KNO₃	0.33	g	0.33	3.264
NaCl	25.00	g	25	427.789
PIPES (12mM)	3.60	g	3.6	11.906
Yeast extract	0.50	g	0.5	-
Tryptone	1.00	g	1	14.069
Sucrose	1.00	g	1	2.921
Modified Wolin's mineral solution II	1.00	ml	-	-
Wolin's vitamin solution	1.00	ml	-	-
Distilled water	1000.00	ml	-	-
1 Prepare the medium anaerobically, under nitrogen, omitting the CaCl₂, MgCl₂, KNO₃, tryptone, yeast extract, vitamins and sucrose. The pH should be 6.8. Dispense the medium into vessels suitable for anaerobic growth (Hungate tubes or serum bottles) under an atmosphere of nitrogen and autoclave. To the sterile, cooled medium add, from sterile stock solutions the CaCl₂, MgCl₂, KNO₃, tryptone, yeast extract, vitamins and sucrose. The CaCl₂, MgCl₂, KNO₃, tryptone, yeast extract, and sucrose stock solutions should be autoclaved, while the vitamin solution is sterile filtered. This strain grows anaerobically in the presence of nitrate, but aerobic growth has been reported provided the oxygen concentration does not exceed 16% (v/v). Under anaerobic conditions a redox agent, such as sulphide is not required.

977	VULCANITHERMUS MEDIUM