DSMZ 716 . AMMONIFEX DEGENSII MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 x 3 H2O 0.30 g 0.299 1.717
KH2PO4 0.22 g 0.219 1.612
(NH4)2SO4 0.22 g 0.219 1.66
NaCl 1.00 g 0.997 17.06
MgSO4 x 7 H2O 0.09 g 0.09 0.364
CaCl2 x 2 H2O 0.06 g 0.06 0.407
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
2.00 ml 0.002 0.007
NiCl2 x 6 H2O
(0.1% w/v)
0.20 ml 2.0e-4 0.002
Modified Wolin's mineral solution 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
KNO3 1.00 g 0.997 9.861
Na2CO3 1.50 g 1.496 14.11
Na2S x 9 H2O 0.30 g 0.299 1.245
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except nitrate, carbonate and sulfide), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic serum vials (e.g., 10 ml medium in 25 ml Balch-type tubes) and autoclave. Prior to use add nitrate from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Before inoculation add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH of the medium to 7.2.
2 After inoculation pressurize vials to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Solution occurs in 166 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 7.2
Equipment needed: Autoclave, Gassing station
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Gas composition

  • 80 % H2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 1.496 14.11
KNO3 0.997 9.861
K2HPO4 x 3 H2O 0.299 1.717
Na2S x 9 H2O 0.299 1.245
(NH4)2SO4 0.219 1.66
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain