# J950: NH-JN4 MEDIUM


## Main sol. J950

1	g	Na2SO4	
30	g	NaCl	
0.7	g	KCl	
5	g	MgCl2 x 6 H2O	
1	g	CaCl2 x 2 H2O	
0.4	g	K2HPO4	
0.4	g	KH2PO4	
0.5	g	NH4Cl	
2	mg	Fe(NH4)2(SO4)2 x 6 H2O	
3	g	Yeast extract (BD-Difco)	
4	g	Tryptone (BD-Difco)	
6.8	g	Sodium acetate	
0.6	g	L-Cysteine HCl x H2O	
1	ml	**Trace element solution SL-10**	
1	mg	Resazurin	
960	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 6.8. Distribute the medium into culture vessels under a N2-CO2 (9:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add the following solution (per 960 ml) from anaerobic stock (filter-sterilized) 

40	ml	NaHCO3 (5%)	


## Trace element solution SL-10

10	ml	HCl (25%, 7.7 M)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts.