# 897: IGNICOCCUS MEDIUM


## Main sol. 897

13.65	g	NaCl	
3.5	g	MgSO4 x 7 H2O	
2.75	g	MgCl2 x 6 H2O	
0.5	g	KH2PO4	
0.25	g	(NH4)2SO4	
0.38	g	CaCl2 x 2 H2O	
0.33	g	KCl	
0.05	g	NaBr	
15	mg	H3BO3	
7	ml	SrCl2 x 6 H2O (0.1% w/v)	
0.5	ml	KI (0.01% w/v)	
0.5	ml	Sodium resazurin (0.1% w/v)	
5	g	Sulfur (powdered)	
1	g	Meat extract	
0.2	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except sulfur, meat extract and sulfide, adjust pH to 5.0 - 5.5 with 2 N H2SO4 and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials already containing the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. For sterilization heat cultivation vessels in boiling water for 1 -2 hours on each of 3 successive days. Add meat extract and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere.
2. After inoculation pressurize the vessels with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.