320
320 .
CLOSTRIDIUM CELLULOVORANS MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Main sol. 320 #
ml
Equipment needed: Autoclave
Filter
Gassing station
Hungate tubes
Centrifuge
| 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). | |||||
Solution occurs in 265
other media (first defined in Medium 320).
| 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. | |||||
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: | 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 | |
Gas composition
- 80 % N2
- 20 % 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: | 1 |
Molecular composition
| 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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