666b
666b .
METHANOMICROBIUM MOBILE MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Main sol. 666b #
ml
Equipment needed: Autoclave
Hungate tubes
Gassing station
Centrifuge
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Clarified rumen fluid | 400.00 | ml | - | - | |
| K2HPO4 | 0.23 | g | 0.23 | 1.321 | |
| KH2PO4 | 0.23 | g | 0.23 | 1.69 | |
| NaCl | 0.45 | g | 0.45 | 7.7 | |
| (NH4)2SO4 | 0.45 | g | 0.45 | 3.405 | |
| CaCl2 x 2 H2O | 0.06 | g | 0.06 | 0.408 | |
| MgSO4 x 7 H2O | 0.09 | g | 0.09 | 0.365 | |
| Indigocarmine | 5.00 | mg | 0.005 | 0.011 | |
| NaHCO3 | 6.40 | g | 6.4 | 76.184 | |
| Yeast extract | 5.00 | g | 5 | - | |
| DL-Dithiothreitol | 0.30 | g | 0.3 | 1.944 | |
| Na2S x 9 H2O | 0.30 | g | 0.3 | 1.249 | |
| Distilled water | 600.00 | ml | - | - | |
| 1 Dissolve ingredients (except bicarbonate, cysteine, and DTT), bring the medium to a boil, and then cool it to room temperature under an 80% H2 and 20% CO2 gas atmosphere. Add the bicarbonate and equilibrate the medium to pH 6.8. Distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials, filling them to 30% of their volume, and then autoclave. After that, add cysteine and DTT from sterile anoxic stock solutions prepared under a 100% N2 gas atmosphere. Adjust the pH of the complete medium to 6.7 - 6.8, if necessary. | |||||
| 2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 2 bar of overpressure. | |||||
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. |
Metadata
| Last modified: | 19.09.25 |
| Source: | MediaDive: DSMZ |
| Taxonomic range: | Archaea |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | 6.8 |
| Equipment needed: | Autoclave, Centrifuge, Gassing station, Hungate tubes |
| Edit in Medium builder | |
Gas composition
- 80 % H2
- 20 % CO2
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| NaHCO3 | 6.4 | 76.184 |
| Yeast extract | 5 | - |
| NaCl | 0.45 | 7.7 |
| (NH4)2SO4 | 0.45 | 3.405 |
| DL-Dithiothreitol | 0.3 | 1.944 |
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