# J151: PYROCOCCUS MEDIUM


## Main sol. J151

1000	ml	**Salt base solution**	
10	ml	**Trace minerals**	
5	mg	Citric acid	
1	g	Yeast extract (BD-Difco)	
5	g	Bacto peptone (BD-Difco)	
30	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 under a N2 atmosphere. 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. Prior to inoculation, add the sterile Na2S x 9H2O solution, and readjust pH to 6.5, if necessary.


## Salt base solution

13.85	g	NaCl	
3.5	g	MgSO4 x 7 H2O	
2.75	g	MgCl2 x 6 H2O	
0.325	g	KCl	
0.05	g	NaBr	
15	mg	H3BO3	
7.5	mg	SrCl2 x 6 H2O	
0.05	mg	KI	
0.75	g	CaCl2	
0.5	g	KH2PO4	
2	mg	(NH4)2Ni(SO4)2 x 6 H2O	
1000	ml	Distilled water	


## 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.