# 1101: DESULFONATRONOSPIRA MEDIUM


## Main sol. 1101

15	g	NaCl	
0.5	g	K2HPO4	
15	g	NaHCO3	
95	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-L-lactate	
1.3	g	Na2SO3	
1	ml	**Wolin's vitamin solution (10x)**	
0.24	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve sodium chloride and hydrogenphospate, then sparge solution with 100% N2 gas for 30 -45 min to make it anoxic. Add and dissolve carbonates, adjust pH to 10, 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, lactate, sulfite, vitamins, and sulfide from sterile anoxic stock solution prepared under 100% N2 gas. Stock solutions of sulfite (freshly prepared) and vitamins should be sterilized by filtration.


## 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