# J508: MODIFIED MMJS MEDIUM


## Main sol. J508

20	g	NaCl	
0.09	g	K2HPO4	
0.07	g	KH2PO4	
0.8	g	CaCl2 x 2 H2O	
0.25	g	NH4Cl	
1	g	NaNO3	
4	g	MgSO4 x 7 H2O	
3	g	MgCl2 x 6 H2O	
0.33	g	KCl	
1	mg	NiCl2 x 6 H2O	
1	mg	Na2SeO3 x 5 H2O	
0.01	g	Fe2(SO4)3 x n H2O	
1	mg	H2WO4	
2	g	Na2S2O3 x 5 H2O	
3	g	Sulfur (powder)	
10	ml	**Trace mineral solution**	
1	ml	**Trace vitamins**	
2	g	NaHCO3	
1000	ml	Distilled water	

1. Mix ingredients except sulfur, Trace vitamins and NaHCO3, adjust pH to 6.8 and autoclave. Steam sulfur for 3 hr on each of 3 successive days. Filter-sterilize Trace vitamins and 5% NaHCO3 solution, and add to the medium. Distribute the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) containing sulfur under a stream of N2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Replace the gas phase with N2-CO2-O2 (79:20:1, v/v) and pressurized inoculated bottles to 200 kPa with the same gas mixture.


## Trace mineral solution

1. Use Trace mineral solution of Medium No. 413 with 0.01 g/L (final) Na2MoO4 x 2H2O.


## 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