# J782: CALDINITRATIRUPTOR MICROAEROPHILUS MEDIUM


## Main sol. J782

0.3	g	KH2PO4	
0.3	g	K2HPO4	
1	g	NH4Cl	
1.7	g	KNO3	
1	g	NaCl	
0.1	g	KCl	
0.1	g	CaCl2 x 2 H2O	
0.25	g	MgCl2 x 6 H2O	
10	ml	**Trace minerals**	
1	g	Yeast extract	
1000	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 7.2 with KOH and distribute the medium into culture vessels (e.g., 5 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add per liter the following solutions from anaerobic stocks (autoclaved or *filter-sterilized): 

25	ml	NaHCO3 (*, 8%)	
20	ml	Glucose (1 M)	


## Trace minerals

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.1	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.01	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.