140
140 .
RUMINICLOSTRIDIUM MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Main sol. 140 #
ml
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
Centrifuge
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.50 | g | 0.5 | 3.674 | |
| NaCl | 1.00 | g | 1 | 17.112 | |
| (NH4)2SO4 | 0.50 | g | 0.5 | 3.784 | |
| MgSO4 x 7 H2O | 0.10 | g | 0.1 | 0.406 | |
| CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.68 | |
| K2HPO4 | 0.50 | g | 0.5 | 2.871 | |
| Clarified rumen fluid | 300.00 | ml | - | - | |
|
Sludge fluid
, alternative
optional |
300.00 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
| Na2CO3 | 2.50 | g | 2.5 | 23.588 | |
| 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 | 700.00 | ml | - | - | |
| 1 Dissolve ingredients (except carbonate, cellobiose and reducing agents), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Use either clarified rumen fluid or sludge fluid as supplement. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Cellobiose has to be sterilized by filtration. Adjust pH of complete medium to 6.8, if necessary. | |||||
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. | |||||
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: | 24.02.22 |
| Source: | DSMZ |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | 6.8 |
| Equipment needed: | Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes |
| Edit in Medium builder | |
Gas composition
- 100 % 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: | 2 |
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| Cellobiose | 5 | 14.607 |
| Na2CO3 | 2.5 | 23.588 |
| Yeast extract | 1.2 | - |
| NaCl | 1 | 17.112 |
| (NH4)2SO4 | 0.5 | 3.784 |
| Show more | ||
