# 1845: Nitrososphaera gargensis medium


## Main sol. 1845

50	ml	**Basal salt stock solution (20x)**	
1	ml	**Trace element solution (Nitrososphaera)**	
1	ml	**FeNaEDTA solution (7.5 mM)**	
1	ml	**Selenite-tungstate solution**	
0.5	ml	**NH4Cl (1 M)**	

1. Add 50 mL of 20x Basal salt stock solution and 4 g CaCO3 to 950 mL sterile double distilled water and mix all just before use.
2. Autoclave at 121 °C for 20 min.
3. Cool down to room temperature.
4. Add aseptic per liter of 1x basalt salt solution: 1 mL FeNaEDTA (7.5 mM), 1 mL trace elements solution, 1 mL Selenite-tungstate solution, 0.5 mL NH4Cl solution.
5. Adjust pH to 8.5 with NaOH.


## Basal salt stock solution (20x)

1	g	KH2PO4	
1.5	g	KCl	
7	g	Mg(SO4) x 7 H2O	
11.68	g	NaCl	

1. Prepare with double distilled water.
2. Autoclave at 121 °C for 20 min.
3. Cool down to room temperature.
4. Store in the dark at 4 °C.


## Trace element solution (Nitrososphaera)

8	mL	HCl (12.5M)	
30	mg	H3BO3	
100	mg	MnCl2 x 4 H2O	
190	mg	CoCl2 x 6 H2O	
24	mg	NiCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
144	mg	ZnSO4 x 7 H2O	
36	mg	Na2MoO4 x 2 H2O	
1000	ml	Double distilled water	

1. This solution contains no iron! Sterilise by autoclaving or filtration (0.2 µm) and store in the dark at 4 °C.


## FeNaEDTA solution (7.5 mM)

2753	mg	FeNa-EDTA	
1000	ml	Double distilled water	

1. Sterilise by filtration (0.2 µm) and store in the dark at 4 °C.


## Selenite-tungstate solution

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


## NH4Cl (1 M)

2.67	g	NH4Cl	
50	ml	Double distilled water	

1. Dissolve 2.67 g ammonium chloride in 50 mL Double distilled water.
2. Filtrate (0.2 µm).