Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Mineral solution | 38.00 | ml | - | - | |
Clarified rumen fluid | 200.00 | ml | - | - | |
K2HPO4 | 0.30 | g | 0.3 | 1.722 | |
Trypticase peptone
(BD BBL) |
2.00 | g | 2 | - | |
Yeast extract
(OXOID) |
0.50 | g | 0.5 | - | |
Haemin solution
(0.05% w/v) |
2.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
Na2CO3 | 4.00 | g | 4 | 37.74 | |
Cellobiose | 5.00 | g | 5 | 14.607 | |
L-Cysteine HCl x H2O | 0.25 | g | 0.25 | 1.423 | |
Na2S x 9 H2O | 0.25 | g | 0.25 | 1.041 | |
Distilled water | 760.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, cellobiose, cysteine and sulfide), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Add the carbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute under CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Cellobiose has to be sterilized by filtration. Adjust pH of complete medium to 6.7 - 6.8, if necessary. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
KH2PO4 | 6.0 | g | 6 | 44.09 |
NaCl | 12.0 | g | 12 | 205.339 |
(NH4)2SO4 | 6.0 | g | 6 | 45.406 |
CaCl2 x 2 H2O | 1.6 | g | 1.6 | 10.883 |
MgSO4 x 7 H2O | 2.5 | g | 2.5 | 10.143 |
Distilled water | 1000.0 | ml | - | - |
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. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Haemin | 50 | mg | 0.5 | 0.767 | |
NaOH
(1 N) |
1 | ml | - | - | |
Distilled water | 100 | ml | - | - | |
1 Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated. |
Last modified: | 04.04.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 6.7 - 6.8 |
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: | 2 |
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Cellobiose | 5 | 14.607 |
Na2CO3 | 4 | 37.74 |
Trypticase peptone | 2 | - |
Yeast extract | 0.5 | - |
NaCl | 0.456 | 7.803 |
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