# J908: HALORHODOSPIRA NEUTRIPHILA MEDIUM


## Main sol. J908

1	g	KH2PO4	
0.5	g	NH4Cl	
100	g	NaCl	
2	g	MgCl2 x 6 H2O	
0.5	g	MgSO4 x 7 H2O	
0.05	g	CaCl2 x 2 H2O	
1	ml	**Trace element solution SL-12**	
20	ml	NaHCO3 (10%, w/v)	
7.5	ml	Na2S x 9 H2O (10%, w/v)	
1	ml	**Seven vitamins solution**	
4	ml	Sodium acetate (0.5 M)	
2	ml	Sodium succinate (0.5 M)	
10	ml	Yeast extract (5%)	
1000	ml	Distilled water	

1. Adjust pH to 7.0 with 1 M Na2CO3 or 1 M H2SO4. Mix ingredients, except NaHCO3, Na2S x 9H2O, seven vitamins solution, acetate solution, succinate solution and yeast extract solution, and autoclave under a N2-CO2 (80:20, v/v) atmosphere. Separately autoclave acetate solution, succinate solution and yeast extract solution. Filter-sterilize NaHCO3, Na2S x 9H2O. Aseptically and anaerobically add NaHCO3, Na2S x 9H2O, seven vitamins solution, acetate solution, succinate solution and yeast extract solution to the medium.


## Trace element solution SL-12

3	g	Na2-EDTA	
1.1	g	FeSO4 x 7 H2O	
190	mg	CoCl2 x 6 H2O	
50	mg	MnCl2 x 4 H2O	
42	mg	ZnCl2	
24	mg	NiCl2 x 6 H2O	
18	mg	Na2MoO4 x 2 H2O	
300	mg	H3BO3	
2	mg	CuCl2 x 2 H2O	
1000	ml	Distilled water	

1. Autoclave separately.
2. Adjust pH to 6.8.


## Seven vitamins solution

10	mg	Vitamin B12	
10	mg	p-Aminobenzoic acid	
2	mg	D-(+)-biotin	
20	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
50	mg	Pyridoxine hydrochloride	
10	mg	Thiamine-HCl x 2 H2O	
200	ml	Distilled water