Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Casitone
(BD Bacto) |
10.00 | g | 10 | - | |
Yeast extract
(BD Bacto) |
2.50 | g | 2.5 | - | |
D-Glucose | 2.00 | g | 2 | 11.101 | |
Cellobiose | 2.00 | g | 2 | 5.843 | |
Maltose | 2.00 | g | 2 | 5.843 | |
Na-DL-lactate | 5.00 | g | 5 | 44.619 | |
Clarified rumen fluid | 200.00 | ml | - | - | |
K2HPO4 | 0.05 | g | 0.05 | 0.287 | |
KH2PO4 | 0.05 | g | 0.05 | 0.367 | |
(NH4)2SO4 | 0.10 | g | 0.1 | 0.757 | |
NaCl | 0.10 | g | 0.1 | 1.711 | |
CaCl2 x 2 H2O | 0.01 | g | 0.01 | 0.068 | |
MgSO4 x 7 H2O | 0.01 | g | 0.01 | 0.041 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
Na2CO3 | 4.00 | g | 4 | 37.74 | |
L-Cysteine HCl x H2O | 0.30 | g | 0.3 | 1.708 | |
Na2S x 9 H2O | 0.30 | g | 0.3 | 1.249 | |
Distilled water | 800.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, cysteine and sulfide), boil medium for 1 min, then cool to room temperature under 100% CO2 gas atmosphere. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Add cysteine and sulfide form sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of the complete medium to 6.5 - 6.8, if necessary. |
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. |
Last modified: | 04.05.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 6.5 - 6.8 |
Equipment needed: | Autoclave, Centrifuge, Gassing station, Hungate tubes |
Edit in Medium builder |
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
All strains for this medium: | 1 |
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Casitone | 10 | - |
Na-DL-lactate | 5 | 44.619 |
Na2CO3 | 4 | 37.74 |
Yeast extract | 2.5 | - |
D-Glucose | 2 | 11.101 |
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