# J421: MMJHS MEDIUM


## Main sol. J421

1000	ml	**MJ(-N) synthetic seawater**	
1	g	NaNO3	
1	g	Na2S2O3 x 5 H2O	
1	g	NaHCO3	
10	ml	**Trace vitamins**	
3	g	Sulfur (powder)	

1. Mix ingredients except sulfur, adjust pH to 6.5 - 7.0 with NaOH and filter-sterilize. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) containing sulfur under a H2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Pressurize the inoculated bottles to 300 kPa H2-CO2 (4:1, v/v).
2. Comment: Adjust final pH to 7.0 (for strain JCM 12458) or 6.5 (for strain JCM 12459).


## 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