909 .
TREPONEMA DENTICOLA MEDIUM
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* Medium for Brucepastera parasyntrophica DSM 111712
Omit agar from solution A.
Growth conditions: 2-3 days
anaerobic
30 °C
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 1000 | ml | ||
|
|
Solution B | 45 | ml | ||
| 1 Add 0.45 ml Sol. B to 10 ml Sol. A. Inoculate and pressurize the tubes with H2 to 0.5 bar. | |||||
Solution A #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Brain heart infusion broth | 12.50 | g | ||
|
|
Trypticase
(BBL 11921) |
10.00 | g | ||
|
|
Yeast extract | 2.50 | g | ||
|
|
Na-thioglycolate | 0.50 | g | ||
|
|
L-Cysteine HCl | 1.00 | g | ||
|
|
L-Asparagine | 0.25 | g | ||
|
|
Glucose | 2.00 | g | ||
| Agar* | 3.00 | g | |||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Boil for 5 min, cool to 50°C while gassing with 80% N2 + 20% CO2. Distribute to anaerobic tubes under the same atmosphere and autoclave. | |||||
Solution B #
45
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Thiamine pyrophosphate | 6 | mg | ||
|
|
NaHCO3 | 2 | g | ||
|
|
Distilled water | 25 | ml | ||
|
|
Volatile fatty acid solution | 20 | ml | ||
|
|
Rabbit serum | 20 | ml | ||
| 1 Sterilize by filtration, add 20 ml rabbit serum, gas with 80% N2 + 20% CO2. | |||||
Volatile fatty acid solution #
102
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KOH
(0.1 N) |
100.0 | ml |
|
|
Isobutyric acid | 0.5 | ml |
|
|
DL-2-Methylbutyric acid | 0.5 | ml |
|
|
Isovaleric acid | 0.5 | ml |
|
|
Valeric acid | 0.5 | ml |