DSMZ 119b . METHANOMASSILIICOCCUS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Hungate tubes Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.5 3.674
MgSO4 x 7 H2O 0.40 g 0.4 1.623
NaCl 0.40 g 0.4 6.845
NH4Cl 0.40 g 0.4 7.478
CaCl2 x 2 H2O 0.05 g 0.05 0.34
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
FeSO4 x 7 H2O solution (0.1% w/v) 2.00 ml - -
Yeast extract
(OXOID)
1.00 g 1 -
Na-acetate 1.00 g 1 12.19
Na-formate 2.00 g 2 29.409
Sludge fluid 50.00 ml - -
Fatty acid mixture 20.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
Methanol 1.50 ml 1.188 37.076
NaHCO3 1.00 g 1 11.904
L-Cysteine HCl x H2O 0.50 g 0.5 2.847
Na2S x 9 H2O 0.50 g 0.5 2.082
Distilled water 930.00 ml - -
1 Dissolve ingredients except methanol, bicarbonate, cysteine and sulfide. Adjust pH of medium to 7.2 and sparge with 100% N2 gas for 30 - 45 min to make it anoxic. Then dispense medium under same gas atmosphere into anoxic Hungate-type tubes to 30% v/v of their volume and autoclave. Add methanol, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate 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 7.6, if necessary. After inoculation, add sterile 80% H2 and 20% CO2 gas mixture to 0.5 bar overpressure. After growth becomes visible overpressure of 80% H2 and 20% CO2 gas mixture can be increased to 1 bar.
2 Note: Use 10% (v/v) as inoculum.
Solution occurs in 262 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%)
10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
Solution occurs in 173 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
Solution occurs in 8 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.4 g 4 -
Sludge 100.0 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 room temperature in the dark.
Solution occurs in 7 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Isobutyric acid 23 ml - -
DL-2-Methylbutyric acid 27 ml - -
Valeric acid 27 ml - -
Isovaleric acid 27 ml - -
Distilled water 1000 ml - -
1 Adjust pH to 7.0 - 7.3 with 2 N NaOH.
Solution occurs in 6 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 0.1 g 1 3.597
H2SO4
(0.1 N)
100.0 ml - -
1 The ferrous sulfate solution is not stable and should be freshly prepared.

Metadata

Last modified: 22.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.6
Equipment needed: Autoclave, Centrifuge, Gassing station, Hungate tubes
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Gas composition

  • 100 % N2

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 2 29.409
Na-acetate 1 12.19
NaHCO3 1 11.904
KH2PO4 0.5 3.674
Na2S x 9 H2O 0.5 2.082
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain