# 561: SULFURIFERULA MEDIUM


## Main sol. 561

0.3	g	KCl	
2.75	g	MgCl2 x 6 H2O	
3.45	g	MgSO4 x 7 H2O	
1.25	g	NH4Cl	
0.14	g	CaCl2 x 2 H2O	
0.14	g	K2HPO4	
0.5	g	NaCl	
10	ml	**Modified Wolin's mineral solution**	
5	ml	NiCl2 x 6 H2O (0.1% w/v)	
0.5	ml	Na2WO4 x 2 H2O (0.1% w/v)	
1000	ml	Distilled water	

1. Dissolve ingredients, adjust pH to 6.0 with H2SO4 and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Adjust medium pH to 6.0, if necessary.
2. After inoculation, add by injection 3% (v/v) sterile air (usually 9 ml per bottle) and pressurize bottles with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.


## 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.