323 .
TREPONEMA SACCHAROPHILUM MEDIUM
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Selected strain: Treponema saccharophilum DSM 2985
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
37 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % CO2
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Final pH:
7.0
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
CaCl2 x 2 H2O | 0.12 | g | ||
|
|
MgSO4 x 7 H2O | 0.19 | g | ||
|
|
KH2PO4 | 0.45 | g | ||
|
|
K2HPO4 | 0.45 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
(NH4)2SO4 | 0.90 | g | ||
|
|
Yeast extract
(OXOID) |
2.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
2.00 | g | ||
|
|
n-Butyric acid | 0.40 | ml | ||
|
|
iso-Butyric acid | 0.40 | ml | ||
|
|
DL-2-Methylbutyric acid | 0.20 | ml | ||
|
|
n-Valeric acid | 0.20 | ml | ||
|
|
iso-Valeric acid | 0.20 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 1.00 | g | ||
|
|
Na2CO3 | 2.50 | g | ||
|
|
D-Glucose | 2.00 | g | ||
|
|
Agar
, for solid medium
(BD Bacto) |
12.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except cysteine, carbonate and glucose), adjust pH to 7.0 and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the cysteine and carbonate, then equilibrate the medium with the CO2 gas to pH 7.0. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add glucose from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary. | |||||
| 2 For solid medium add 12.00 g/l agar (BD Bacto). | |||||