# J1083: DESULFURELLA TR1 MEDIUM


## Main sol. J1083

0.53	g	Na2HPO4 x 2 H2O	
0.41	g	KH2PO4	
0.3	g	NH4Cl	
0.3	g	NaCl	
0.1	g	MgCl2 x 6 H2O	
0.11	g	CaCl2 x 2 H2O	
1	ml	**Acid trace element solution**	
1	ml	**Alkaline trace element solution**	
0.68	g	Sodium acetate x 3 H2O	
0.1	g	Yeast extract	
0.8	g	Sulfur (powder)	
1	m	Resazurin	
1000	ml	Distilled water	

1. Mix components, except sulfur, and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized): 

0.2	ml	**Vitamin solution**	
12.5	ml	NaHCO3 (8%)	

2. Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically distribute the medium and sulfur into culture vessels (e.g., 50 ml of the medium and 0.04 g of sulfur in 120 ml serum bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, anaerobically add per liter the following solution from an anaerobic stock (autoclaved and stored under a N2 atmosphere): 

6	ml	Na2S x 9 H2O (5%)	

3. Adjust pH to 6.0, if necessary.


## Acid trace element solution

10	ml	HCl (6 N)	
61.8	mg	H3BO3	
61.25	mg	MnCl2 x 4 H2O	
943.5	mg	FeCl2 x 4 H2O	
64.5	mg	CoCl2 x 6 H2O	
12.86	mg	NiCl2 x 6 H2O	
67.7	mg	ZnCl2	
1000	ml	Distilled water	


## Alkaline trace element solution

400	mg	NaOH	
17.3	mg	Na2SeO3 x 5 H2O	
29.4	mg	Na2WO4 x 2 H2O	
20.5	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	


## Vitamin solution

20	mg	Biotin	
200	mg	Nicotinic acid	
100	mg	p-Aminobenzoic acid	
200	mg	Thiamine HCl	
100	mg	DL-Calcium pantothenate	
500	mg	Pyridoxine hydrochloride	
100	mg	Vitamin B12	
100	mg	Riboflavin	
1000	ml	Distilled water	

1. Filter-sterilize and store under a N2 atmosphere.