# J344: HNW MEDIUM


## Main sol. J344

670	ml	**MJ(-N) synthetic seawater**	
0.25	g	NH4Cl	
1	g	NaNO3	
0.1	mg	Na2WO4 x 2 H2O	
330	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 7.0 with NaOH and autoclave. After cooling, add the following solutions (filter-sterilized): 

12.5	ml	NaHCO3 (8%)	
10	ml	**Trace vitamins**	

2. Aseptically distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) under a H2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Pressurize inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).
3. Comment: For JCM 34026, use 1.0 L of MJ(-N) synthetic seawater and omit distilled water.


## MJ(-N) synthetic seawater

30	g	NaCl	
0.14	g	K2HPO4	
0.14	g	CaCl2 x 2 H2O	
3.4	g	MgSO4 x 7 H2O	
4.18	g	MgCl2 x 6 H2O	
0.33	g	KCl	
0.5	mg	NiCl2 x 6 H2O	
0.5	mg	Na2SeO3 x 5 H2O	
0.01	g	Fe(NH4)2(SO4)2 x 6 H2O	
10	ml	**Modified Wolin's mineral solution**	
1000	ml	Distilled water	


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water