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DSMZ

DSMZ 321 . METHANOCORPUSCULUM AGGREGANS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 321 #

ml
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.

Minerals solution #

1000
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 - -

Modified Wolin's mineral solution #

1000
Solution occurs in 167 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.

Clarified rumen fluid #

1000
Solution occurs in 28 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.

Sludge fluid #

1000
Solution occurs in 7 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