159
159 .
TREPONEMA MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Main sol. 159 #
ml
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
Centrifuge
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Clarified rumen fluid | 300.00 | ml | - | - | |
| CaCl2 x 2 H2O | 0.12 | g | 0.12 | 0.816 | |
| MgSO4 x 7 H2O | 0.18 | g | 0.18 | 0.73 | |
| KH2PO4 | 0.45 | g | 0.45 | 3.307 | |
| K2HPO4 | 0.45 | g | 0.45 | 2.584 | |
| NaCl | 0.90 | g | 0.9 | 15.4 | |
| (NH4)2SO4 | 0.90 | g | 0.9 | 6.811 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
| L-Cysteine HCl x H2O | 1.00 | g | 1 | 5.694 | |
| Na2CO3 | 1.50 | g | 1.5 | 14.153 | |
| D-Glucose | 2.00 | g | 2 | 11.101 | |
| Distilled water | 700.00 | ml | - | - | |
| 1 Dissolve ingredients (except cysteine, carbonate and glucose) and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH with KOH to 6.5. Distribute medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Therafter, add glucose from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Final pH of the medium should be 6.7 - 7.0. | |||||
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: | 26.10.22 |
| Source: | DSMZ |
| Synonyms: | TREPONEMA BRYANTII MEDIUM |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | 6.7 - 7.0 |
| 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 .
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| D-Glucose | 2 | 11.101 |
| Na2CO3 | 1.5 | 14.153 |
| L-Cysteine HCl x H2O | 1 | 5.694 |
| NaCl | 0.9 | 15.4 |
| (NH4)2SO4 | 0.9 | 6.811 |
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