# J1239: THERMOPLASMA KAMCHATKENSIS MEDIUM


## Main sol. J1239

3	g	KH2PO4	
1	g	MgSO4 x 7 H2O	
0.25	g	CaCl2 x 2 H2O	
0.2	g	(NH4)2SO4	
0.1	g	Yeast extract	
5	g	Sucrose	
4	g	Sulfur (powder)	
1000	ml	Distilled water	

1. Mix components except sulfur, yeast extract and sucrose, adjust pH to 2.5 with 10 N H2SO4 and autoclave. Separately autoclave yeast extract and sucrose and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Aseptically distribute the medium into culture vessels (e.g., 10 ml in Balch tubes) containing appropriate amount of sulfur, replace gas phase with a N2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers.