# J696: THERMODESULFATATOR ATLANTICUS MEDIUM


## Main sol. J696

20	g	NaCl	
3	g	MgCl2 x 6 H2O	
3	g	Na2SO4	
0.5	g	KCl	
0.3	g	NH4Cl	
0.1	g	KH2PO4	
1.73	g	PIPES	
0.11	g	CaCl2 x 2 H2O	
10	ml	**Trace minerals**	
10	ml	**Trace vitamins**	
0.3	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Mix components except Na2S x 9H2O and adjust pH to 6.7. Distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) under a gas stream of H2-CO2 (4:1, v/v), seal with butyl rubber stoppers and autoclave. Separately autoclave 3% Na2S x 9H2O solution under a N2 gas atmosphere. Prior to inoculation, reduce the medium by the addition of the Na2S x 9H2O solution. Pressurized the inoculated culture vessels to 200 kPa H2-CO2 (4:1, v/v).


## 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.


## 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