# 399: ARCHAEOGLOBUS MEDIUM


## Main sol. 399

0.34	g	KCl	
4	g	MgCl2 x 6 H2O	
3.45	g	MgSO4 x 7 H2O	
0.25	g	NH4Cl	
0.14	g	CaCl2 x 2 H2O	
0.14	g	K2HPO4	
18	g	NaCl	
0.5	g	Yeast extract (OXOID)	
1.5	g	Na-L-lactate	
2	ml	Fe(NH4)2(SO4)2 x 7 H2O (0.1% w/v)	
10	ml	**Modified Wolin's mineral solution**	
0.5	ml	Sodium resazurin (0.1% w/v)	
3	g	NaHCO3	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except bicarbonate and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and adjust pH to 6.9, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into serum vials (e.g., 20 ml per 100 ml bottle), seal and autoclave. Before use reduce the medium with sodium sulfide from a sterile anoxic stock solution prepared under 100% N2 gas.
2. After inoculation, add 2 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.


## Modified Wolin's mineral solution

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	
0.03	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
0.4	mg	Na2WO4 x 2 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.