# 783: HYDROGENOTHERMUS HIRSCHII MEDIUM


## Main sol. 783

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	
0.15	g	K2HPO4	
0.5	g	CaCl2 x 2 H2O	
10	ml	**Modified Wolin's mineral solution II**	
0.5	g	Sulfur (powdered)	
1000	ml	Distilled water	

1. Prepare the medium aerobically and adjust pH to 7.0 using H2SO4. Fill 20 ml medium into 100 ml serum bottles under H2/CO2 (80/20) gas and seal with a rubber stopper. Steam medium for 3 hours on each of 3 successive days. Before use add 100 µl of a sterile CaCO3 solution (10%). Give an atmosphere of H2/CO2 with an overpressure of two atmospheres and 20 ml filter-sterilized air. Incubate under shaking at 60°C.


## Modified Wolin's mineral solution II

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.025	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.