# 1020: SULFURICURVUM (MBM) MEDIUM


## Main sol. 1020

0.2	g	NaNO3	
0.2	g	KH2PO4	
0.2	g	NH4Cl	
0.4	g	MgCl2 x 6 H2O	
0.2	g	KCl	
0.1	g	CaCl2 x 2 H2O	
2	ml	**Trace element solution SL-4**	
2.5	g	Na2S2O3 x 5 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients, except sodium thiosulfate, then sparge solution with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 7.0, then distribute medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Prior to inoculation, add thiosulfate from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
2. After inoculation, pressurize culture vessels to 0.5 bar overpressure with sterile 100% H2 gas.


## Trace element solution SL-4

0.5	g	Na2-EDTA	
0.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds.