# 1166a: DESULFURIBACILLUS MEDIUM (ARSENATE)


## Main sol. 1166a

6	g	NaCl	
1	g	K2HPO4	
2.8	g	Na2SO4	
8	g	NaHCO3	
22	g	Na2CO3	
0.2	g	NH4Cl	
0.2	g	MgCl2 x 6 H2O	
1	ml	**Trace elements solution (Pfennig & Lippert,1966)**	
1	ml	**Selenite-tungstate solution**	
0.05	g	Yeast extract	
2.2	g	Na-pyruvate	
6.2	g	Na2HAsO4 x 7 H2O	
1	ml	**Wolin's vitamin solution (10x)**	
0.24	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve sodium chloride, hydrogenphosphate and sodium sulfate, then sparge solution with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve hydrogencarbonate and carbonate, then dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add ammonium chloride, magnesium chloride, trace elements, yeast extract, pyruvate, arsenate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of vitamins, pyruvate and arsenate should be sterilized by filtration. Adjust pH of complete medium to 9.5 - 10.


## Trace elements solution (Pfennig & Lippert,1966)

5	g	EDTA	
2.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.03	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.03	g	CuCl2 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. pH 3.0-4.0


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water