# J347: SME MEDIUM


## Main sol. J347

500	ml	**Synthetic seawater (2 x)** (2 x)	
1	g	KH2PO4	
0.5	g	(NH4)2SO4	
0.4	g	Yeast extract (BD-Difco)	
10	g	Sulfur (powder)	
1	mg	Resazurin	
0.5	g	Na2S x 9 H2O	
500	ml	Distilled water	

1. Mix ingredients, except yeast extract, sulfur and Na2S x 9H2O, and autoclave. Separately autoclave 10% (w/v) yeast extract solution, and 5% Na2S x 9H2O solution under a N2 atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) containing sulfur under a stream of H2-CO2 (4:1, v/v) gas mixture, add the yeast extract solution, and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S x 9H2O solution, and adjust pH to 5.5 with sterile 1 N H2SO4. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v) gas mixture.


## Synthetic seawater (2 x)

55.4	g	NaCl	
14	g	MgSO4 x 7 H2O	
11	g	MgCl2 x 6 H2O	
1.3	g	KCl	
0.2	g	NaBr	
0.06	g	H3BO3	
0.03	g	SrCl2 x 6 H2O	
0.1	mg	KI	
20	mg	Trisodium citrate	
1.5	g	CaCl2 x 2 H2O	
1000	ml	Distilled water