# 1101a: DESULFONATRONUM ZHILINAE MEDIUM


## Main sol. 1101a

25	g	NaCl	
0.5	g	K2HPO4	
0.2	g	Yeast extract	
3	g	Na2SO4	
1.85	g	NaHCO3	
1.2	g	Na2CO3	
0.1	g	MgCl2 x 6 H2O	
1	ml	**Trace elements solution (Pfennig & Lippert,1966)**	
1	ml	**Selenite-tungstate solution**	
2.2	g	Na-L-lactate	
1	ml	**Seven vitamins solution**	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except carbonates, magnesium chloride, trace elements, lactate, vitamins and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Then add and dissolve carbonates, adjust pH to 9.0, dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add magnesium chloride, trace elements, lactate, vitamins and sulfide from sterile anoxic stock solution prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Adjust pH of complete medium to 9.0 - 9.5, if necessary.


## 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	


## Seven vitamins solution

100	mg	Vitamin B12	
80	mg	p-Aminobenzoic acid	
20	mg	D-(+)-biotin	
200	mg	Nicotinic acid	
100	mg	Calcium pantothenate	
300	mg	Pyridoxine hydrochloride	
200	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water