# J1290: NRmbB1 MEDIUM


## Main sol. J1290

0.54	g	NH4Cl	
0.14	g	KH2PO4	
3	g	MgCl2 x 6 H2O	
0.15	g	CaCl2 x 2 H2O	
20	g	NaCl	
1	ml	**Trace mineral solution**	
1	ml	**Se/W solution**	
2.5	g	NaHCO3	
1	mg	Resazurin	
900	ml	Distilled water	

1. Mix components except NaHCO3 and adjust pH to 6.5. Bring to a boil for 5-10 sec, then, cool down under a stream of N2-CO2 (80:20, v/v) and add NaHCO3 to the solution. Dispense the medium into culture vessels under a stream of N2-CO2 (80:20, v/v), seal with butyl rubber stopper and autoclave. After autoclaving, stand the medium overnight. Aseptically and anaerobically add per liter the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized): 

1	ml	Vitamin solution (see Medium No. 197, *)	
0.1	ml	**Vitamin B12 solution 0.05 g/L** (*)	
20	ml	Glucose (1.0 M)	
10	ml	Coenzyme M (0.5 M)	
6	ml	Na2S x 9 H2O (5%)	


## Trace mineral solution

1.99	g	FeCl2 x 4 H2O	
0.198	g	MnCl2 x 4 H2O	
0.13	g	CoCl2 x 6 H2O	
0.136	g	ZnCl2	
1.7	mg	CuCl2 x 2 H2O	
6.2	mg	H3BO3	
0.024	g	Na2MoO4 x 2 H2O	
0.013	g	NiCl2 x 6 H2O	
0.013	g	AlCl3	
1000	ml	Distilled water	


## Se/W solution

1.7	mg	Na2SeO3 x 5 H2O	
3.3	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Vitamin B12 solution 0.05 g/L

5	mg	Vitamin B12	
100	ml	Distilled water