# J753: DEHALOSPIRILLUM MEDIUM


## Main sol. J753


## Solution A

0.7	g	Na2SO4	
0.2	g	K2HPO4	
0.25	g	NH4Cl	
0.25	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.5	g	KCl	
0.15	g	CaCl2 x 2 H2O	
0.5	mg	Resazurin	
2	g	Yeast extract	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
1	ml	**Selenite-tungstate solution**	
0.05	g	L-Cysteine HCl x H2O	
0.025	g	FeSO4 x 7 H2O	
900	ml	Distilled water	

1. Mix components except L-cysteine x HCl x H2O and FeSO4 x 7H2O. Bring to a boil for a few seconds and cool down under a N2 atmosphere. While gassing with N2, add L-cysteine x HCl x H2O and FeSO4 x 7H2O to the solution, distribute into culture vessels (e.g., 18 ml in 50 ml serum bottles) under a N2 atmosphere, then, seal butyl rubber stoppers and autoclave. Aseptically and anaerobically add per liter (final) the following sterile solutions (autoclaved or *filter-sterilized) from anaerobic stocks: 

10	ml	Potassium phosphate buffer (0.1 M),  pH 7.5	
10	ml	Trace vitamins (see Medium No. 197) (*)	
1	ml	Vitamin B12 solution (see Medium No. 403) (*)	
42	ml	NaHCO3 (*, 8%)	
18	ml	Sodium pyruvate (*, 25%)	
40	ml	Sodium fumarate (*, 16%)	

2. Adjust pH of the medium to 7.3 - 7.7, if necessary.


## FeCl2 solution

10	ml	HCl (7.7 M, 25%)	
1.5	g	FeCl2 x 4 H2O	
990	ml	Distilled water	


## Trace element solution

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	
1000	ml	Distilled water	


## Selenite-tungstate solution

0.4	g	NaOH	
6	mg	Na2SeO3 x 5 H2O	
8	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water