# J206: JCM MEDIUM No. 206


## Main sol. J206

1000	ml	**Salt base solution**	
10	ml	**Trace minerals**	
5	mg	Citric acid	
6	g	Tryptone (BD-Difco)	
6	g	Sulfur (powder)	
0.5	g	Na2S x 9 H2O	
1	mg	Resazurin	

1. Mix ingredients, except sulfur and Na2S x 9H2O, and adjust pH to 6.5 with H2SO4. Sterilize by filtration while gassing with H2-CO2 (80:20) gas mixture. Steam sulfur for 3 hr on each of 3 successive days. Neutralize Na2S x 9H2O as a 5% solution, and autoclave under a N2 atmosphere. Aseptically and anaerobically add sterile sulfur to the medium under a H2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution. Pressurize inoculated bottles to 100 kPa H2-CO2 (80:20).


## Salt base solution

1. Use Salt base solution of Medium No. 151 with 17.0 g/L (final) NaCl and 2.5 mg/L (final) KI.


## Trace minerals

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.1	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.01	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.