# J417: MJYTFSH MEDIUM


## Main sol. J417

20	g	NaCl	
0.9	g	K2HPO4	
0.7	g	KH2PO4	
1	g	CaCl2 x 2 H2O	
1.25	g	NH4Cl	
3.4	g	MgSO4 x 7 H2O	
4.18	g	MgCl2 x 6 H2O	
0.33	g	KCl	
5	mg	NiCl2 x 6 H2O	
5	mg	Na2SeO3 x 5 H2O	
0.01	g	Fe2(SO4)3 x n H2O	
5	mg	H2WO4	
0.02	g	FeSO4 x 7 H2O	
1	g	Yeast extract (BD-Difco)	
1	g	Tryptone (BD-Difco)	
0.2	g	Sodium formate	
10	ml	**Trace mineral solution**	
1	ml	**Trace vitamins**	
2	g	NaHCO3	
10	g	Sulfur (powder)	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Mix ingredients, except Trace vitamins, NaHCO3 and sulfur, adjust pH to 5.5, and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution, and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S x 9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) containing sterilized sulfur under a stream of H2-CO2 (4:1, v/v) gas mixture, sparge for 5 min with the same gas mixture, and seal with butyl rubber stoppers. Prior to inoculation, add the sterilized Na2S x 9H2O solution to the medium, and pressurize to 200 kPa H2-CO2 (4:1, v/v). Readjust pH to 5.5, if necessary.


## Trace mineral solution

1. Use Trace mineral solution of Medium No. 324 supplemented with 0.01 g/L SrCl2 x H2O, 0.01 g/L NaBr and 0.01 g/L KI.


## 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