# J354: MODIFIED SULFATE REDUCING BACTERIA MEDIUM


## Main sol. J354

20	g	NaCl	
3	g	MgCl2 x 6 H2O	
4	g	Na2SO4	
0.2	g	KH2PO4	
0.5	g	KCl	
0.25	g	NH4Cl	
3.46	g	PIPES	
0.15	g	CaCl2 x 2 H2O	
0.5	g	Tryptone (BD-Difco)	
0.166	g	Sodium acetate	
2	mg	Na2WO4 x 2 H2O	
0.5	mg	Na2SeO4	
10	ml	**Trace vitamins**	
0.05	mg	Vitamin B12	
1	mg	Resazurin	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Mix ingredients, except Na2S x 9H2O, and adjust pH to 6.7 with HCl. After autoclaving under a N2 atmosphere, distribute the medium into sterile culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Neutralize 5% Na2S x 9H2O solution, and autoclave under a N2 atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution to the medium. Pressurize the inoculated vessels to 200 kPa H2-CO2 (4:1, v/v).


## 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