# J828: MODIFIED NMS MEDIUM


## Main sol. J828

1	g	MgSO4 x 7 H2O	
0.2	g	CaCl2 x 2 H2O	
1	g	KNO3	
5	ml	**Trace element solution SL-4**	
10	ml	**Chelated iron solution**	
900	ml	Distilled water	

1. Mix components and autoclave. After cooling, add the following solutions (autoclaved) to the medium: 

100	ml	**Phosphate buffer solution**	

2. Adjust pH to 6.8, if necessary. Aseptically distribute the medium into sterile serum bottles (leave &; 80% gas phase by volume) and seal with rubber stoppers. Replace the gas phase with an air-methane gas mixture (75:25, v/v). Cultivate with shaking.


## Chelated iron solution

0.027	g	FeCl3 x 6 H2O	
0.0372	g	Na2-EDTA	
100	ml	Distilled water	


## Phosphate buffer solution

0.272	g	KH2PO4	
0.717	g	Na2HPO4 x 2 H2O	
100	ml	Distilled water	


## Trace element solution SL-4

0.5	g	EDTA	
0.2	g	FeSO4 x 7 H2O	
100	ml	**Trace element solution SL-6**	
900	ml	Distilled water	


## Trace element solution SL-6

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