# 1630b: NITROSOPUMILUS UREIPHILUS MEDIUM


## Main sol. 1630b

26	g	NaCl	
5	g	MgSO4 x 7 H2O	
5	g	MgCl2 x 6 H2O	
1.5	g	CaCl2 x 2 H2O	
0.1	g	KBr	
2	ml	KH2PO4 (0.10% w/v)	
0.25	g	NaHCO3	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
0.05	g	NH4Cl	
0.05	g	Na-pyruvate	
10	ml	**Wolin's vitamin solution**	
2.5	ml	Catalase (SIGMA C1345) (0.10% w/v)	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate, vitamins, and catalase) and dispense medium in Erlenmeyer flasks with screw caps (e.g., from SCHOTT), then autoclave. Add bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate and vitamins from stock solutions sterilized by filtration. Adjust pH of the complete medium to 6.8 using a sterile solution of sodium bicarbonate (10% w/v) or 1 N HCl.
2. After inoculation, add to the medium 5 - 10 units/ml catalase from a freshly prepared 1.00 mg/ml stock solution sterilized by filtration.
3. Note: Use at least 5 - 10% (v/v) inoculum and incubate in the dark without shaking.


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## 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

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 D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water