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DSMZ

DSMZ 1630c . modified Nitrosospaera medium

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Equipment needed: Autoclave Filter
Final pH: 7.5
Description: based on M. 1630, but modified after Melcher&Schleper during deposition in 2021-2022
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Main sol. 1630c #

ml
Compound Amount Unit
NaCl 1.00 g
MgCl2 x 6 H2O 0.40 g
CaCl2 x 2 H2O 0.10 g
KH2PO4 0.20 g
KCl 0.50 g
Na2SO4 0.05 g
Trace element solution (Nitrososphaera) 1.00 ml
FeNaEDTA solution (7.5 mM) 1.00 ml
NaHCO3 buffer (1M) 2.00 ml
HEPES buffer solution 10.00 ml
NH4Cl (1 M) 2.00 ml
Sodiumpyruvate (1 M) 1.00 ml
Double distilled water 1000.00 ml
1 Incubate all used glass ware and magnetic stirring rods in 0.1 M HCl and rinse 3x with double distilled water to remove residual detergents (always check visual for soap bubbles).
2 Autoclave the Basic-FWM and cool to room temperature. This medium should be free of any precipitations!
3 Aseptically, add per L of BASIC-FWM the following solutions from sterile stocks (do not store complemented FWM): - Trace element solution (Nitrososphaera) - FeNaEDTA solution - NaHCO3 solution - HEPES buffer solution - NH4Cl solution - Sodiumpyruvate solution
4 Adjust pH to 7.5.

Trace element solution (Nitrososphaera) #

1000
Compound Amount Unit
HCl
(12.5M) 		8 mL
H3BO3 30 mg
MnCl2 x 4 H2O 100 mg
CoCl2 x 6 H2O 190 mg
NiCl2 x 6 H2O 24 mg
CuCl2 x 2 H2O 2 mg
ZnSO4 x 7 H2O 144 mg
Na2MoO4 x 2 H2O 36 mg
Double distilled water 1000 ml
1 This solution contains no iron! Sterilise by autoclaving or filtration (0.2 µm) and store in the dark at 4 °C.

HEPES buffer solution #

1000
Compound Amount Unit
NaOH
(pellets) 		24.0 g
HEPES
(free acid) 		238.4 g
Distilled water 1000.0 ml
1 Fist dissolve NaOH in ca 600 ml water, then add HEPES, adjust pH to 7.5 and fill up to 1000 ml volume.

FeNaEDTA solution (7.5 mM) #

1000
Compound Amount Unit
FeNa-EDTA 2753 mg
Double distilled water 1000 ml
1 Sterilise by filtration (0.2 µm) and store in the dark at 4 °C.

NaHCO3 buffer (1M) #

50
Compound Amount Unit
NaHCO3 4.2 g
Double distilled water 50.0 ml
1 Dissolve 4.2 g of NaHCO3 in 50 mL Double distilled water (use a small beaker with stirring rod to dissolve) and sterilize by filtration (0.2 µm) in the laminar flow hood. Store in a sterile 50 mL falcon tube at 4 °C. Avoid high headspace to liquid ratios to ensure stable concentration.

NH4Cl (1 M) #

50
Compound Amount Unit
NH4Cl 2.67 g
Double distilled water 50.00 ml
1 Dissolve 2.67 g ammonium chloride in 50 mL Double distilled water.
2 Filtrate (0.2 µm).

Sodiumpyruvate (1 M) #

50
Compound Amount Unit
Sodium pyruvate 5.5 g
Double distilled water 50.0 ml
1 Dissolve 5.5 g in 50 mL and filter it through 0.2 µm sterile filter in a sterile falcon tube. Store at 4 °C