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DSMZ

DSMZ 1630c . modified Nitrosospaera medium

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×

Main sol. 1630c #

ml
Equipment needed: Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 1 17.112
MgCl2 x 6 H2O 0.40 g 0.4 1.968
CaCl2 x 2 H2O 0.10 g 0.1 0.68
KH2PO4 0.20 g 0.2 1.47
KCl 0.50 g 0.5 6.707
Na2SO4 0.05 g 0.05 0.352
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
Solution occurs in 3 other media (first defined in Medium 1630c).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(12.5M) 		8 mL 3.646 100
H3BO3 30 mg 0.03 0.485
MnCl2 x 4 H2O 100 mg 0.1 0.505
CoCl2 x 6 H2O 190 mg 0.19 1.463
NiCl2 x 6 H2O 24 mg 0.024 0.185
CuCl2 x 2 H2O 2 mg 0.002 0.012
ZnSO4 x 7 H2O 144 mg 0.144 0.501
Na2MoO4 x 2 H2O 36 mg 0.036 0.149
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
Solution occurs in 3 other media (first defined in Medium 1630).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH
(pellets) 		24.0 g 24 600.045
HEPES
(free acid) 		238.4 g 238.4 1000.42
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
Solution occurs in 3 other media (first defined in Medium 1630c).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeNa-EDTA 2753 mg 2.753 -
Double distilled water 1000 ml - -
1 Sterilise by filtration (0.2 µm) and store in the dark at 4 °C.

NaHCO3 buffer (1M) #

50
Solution occurs in 2 other media (first defined in Medium 1630c).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 4.2 g 84 999.917
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
Solution occurs in 3 other media (first defined in Medium 1630c).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 2.67 g 53.4 998.317
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
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sodium pyruvate 5.5 g 110 999.636
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

Metadata

Last modified: 17.09.24
Source: MediaDive: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 7.5
Equipment needed: Autoclave, Filter
Description:

based on M. 1630, but modified after Melcher&Schleper during deposition in 2021-2022
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
HEPES 2.384 10.004
NaCl 1 17.112
KCl 0.5 6.707
MgCl2 x 6 H2O 0.4 1.968
NaOH 0.24 6
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
temperature thermophilic 1 strain