# J279: HYDROGENOPHILUS HIRSCHII MEDIUM


## Main sol. J279

7	g	MgSO4 x 7 H2O	
5.5	g	MgCl2 x 6 H2O	
0.65	g	KCl	
0.1	g	NaBr	
2	g	NaHCO3	
0.15	g	NH4Cl	
0.5	g	CaCl2 x 2 H2O	
10	ml	**Trace minerals**	
0.02	g	(NH4)2Ni(SO4)2 x 6 H2O	
0.1	mg	Na2WO4 x 2 H2O	
0.1	mg	Na2SeO4	
1	g	Na2S2O3 x 5 H2O	
1000	ml	Distilled water	

1. Adjust pH to 7.0 with H2SO4. Distribute 20 ml of medium into 120 ml serum bottles under a stream of a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Autoclave and stand the bottles overnight. Add 15 ml filter-sterilized air and pressurize the gas phase to 200 kPa with a H2-CO2 (4:1, v/v) gas mixture.


## 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.