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DSMZ

DSMZ 119b . METHANOMASSILIICOCCUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 119b #

ml
Equipment needed: Autoclave Hungate tubes Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.498 3.66
MgSO4 x 7 H2O 0.40 g 0.398 1.616
NaCl 0.40 g 0.398 6.817
NH4Cl 0.40 g 0.398 7.448
CaCl2 x 2 H2O 0.05 g 0.05 0.339
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 0.996 -
Na-acetate 1.00 g 0.996 12.142
Na-formate 2.00 g 1.992 29.292
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
(15% v/v) 		10.00 ml 7.888 246.191
NaHCO3 1.00 g 0.996 11.856
L-Cysteine HCl x H2O 0.50 g 0.498 2.835
Na2S x 9 H2O 0.50 g 0.498 2.073
Distilled water 940.00 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 (from 15% v/v stock solution), 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.
2 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.
3 Note: Use 10% (v/v) as inoculum.

Trace element solution SL-10 #

1000
Solution occurs in 265 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.

Selenite-tungstate solution #

1000
Solution occurs in 178 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 - -

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.

Fatty acid mixture #

1000
Solution occurs in 6 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 896 ml - -
1 Adjust pH to 7.5 with concentrated NaOH.

FeSO4 x 7 H2O solution (0.1% w/v) #

1000
Solution occurs in 13 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 1 g 1 3.597
H2SO4
(0.1 N) 		1000 ml - -
1 The ferrous sulfate solution is not stable and should be freshly prepared.

Metadata

Last modified: 11.01.24
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

  • 66 % N2
  • 27 % H2
  • 7 % 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]
Methanol 7.888 246.191
Na-formate 1.992 29.292
Na-acetate 0.996 12.142
NaHCO3 0.996 11.856
KH2PO4 0.498 3.66
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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