Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
K2HPO4 x 3 H2O | 1.00 | g | 1 | 5.741 | |
NH4Cl | 1.00 | g | 1 | 18.695 | |
KCl | 0.50 | g | 0.5 | 6.707 | |
MgSO4 x 7 H2O | 0.50 | g | 0.5 | 2.029 | |
Trypticase peptone
(BD BBL) |
0.50 | g | 0.5 | - | |
Yeast extract | 0.50 | g | 0.5 | - | |
Clarified rumen fluid | 20.00 | ml | - | - | |
Sludge fluid
, alternative
optional |
20.00 | ml | - | - | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
L-Cysteine HCl x H2O | 0.15 | g | 0.15 | 0.854 | |
Na2CO3 | 1.00 | g | 1 | 9.435 | |
Cellobiose | 5.00 | g | 5 | 14.607 | |
Cellulose
, MN 301
optional |
10.00 | g | - | - | |
Na2S x 9 H2O | 0.15 | g | 0.15 | 0.625 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except cysteine, carbonate, cellobiose and sulfide), bring medium to the boil, then cool to room temperature under 80% N2 and 20% CO2 gas mixture and add cysteine. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Sterilize cellobiose separately by filtration under 100% N2 gas. Adjust pH of the complete medium to 7.0, if necessary. | |||||
2 Note: Some strains can be adapted to cellulose as substrate using 10.0 g/l cellulose powder MN 301 (MACHEREY-NAGEL). |
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. |
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. |
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: | 31.03.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 7.0 |
Equipment needed: | Autoclave, Centrifuge, Filter, 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] |
---|---|---|
Cellobiose | 5 | 14.607 |
NH4Cl | 1 | 18.695 |
Na2CO3 | 1 | 9.435 |
K2HPO4 x 3 H2O | 1 | 5.741 |
KCl | 0.5 | 6.707 |
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