DSMZ 321 . METHANOCORPUSCULUM AGGREGANS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Autoclave Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 1 18.695
K2HPO4 x 3 H2O 0.4 g 0.4 2.297
MgCl2 x 6 H2O 0.4 g 0.4 1.968
Minerals solution 50.0 ml - -
Na-formate 5.0 g 5 73.522
Na-acetate 1.0 g 1 12.19
Trypticase peptone
(BD BBL)
1.0 g 1 -
Yeast extract
(OXOID)
1.0 g 1 -
Modified Wolin's mineral solution 10.0 ml - -
Sludge fluid 5.0 ml - -
Clarified rumen fluid , alternative
optional
5.0 ml - -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
Na2CO3 1.5 g 1.5 14.153
L-Cysteine HCl x H2O 0.2 g 0.2 1.139
Na2S x 9 H2O 0.2 g 0.2 0.833
Distilled water 935.0 ml - -
1 Dissolve ingredients except carbonate, cysteine 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 Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary.
2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.
Solution occurs in 2 other media (first defined in Medium 317).
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 6.00 g 6 44.09
(NH4)2SO4 6.00 g 6 45.406
NaCl 12.00 g 12 205.339
MgSO4 x 7 H2O 2.60 g 2.6 10.549
CaCl2 x 2 H2O 0.16 g 0.16 1.088
Distilled water 1000.00 ml - -
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.
Solution occurs in 27 other media (first defined in Medium 1310).
1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.
Solution occurs in 8 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 4 g 4 -
Sludge 1000 ml - -
1 Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours. Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at 8-12°C in the dark.

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Centrifuge, Gassing station, Hungate tubes
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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]
Na-formate 5 73.522
Na2CO3 1.5 14.153
NH4Cl 1 18.695
Na-acetate 1 12.19
Trypticase peptone 1 -
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain