144 .
MODIFIED NOS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| 1 Combine the above ingredients and bring to a boil in a round bottom flask while flushing with O2-free N2-H2-CO2 (85:10:5) gas mixture. Stopper anaerobically, wire or seal closed and autoclave at 121C for 20 min. After autoclaving, allow to cool, open without admitting oxygen and while flushing with gas, add the following ingredients which have been filter-sterilized ahead of time: | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Heart Infusion Broth
(BD-Difco) |
1.25 | g | ||
|
|
Trypticase peptone
(BD-BBL) |
1.00 | g | ||
|
|
Yeast extract
(BD-BBL) |
0.25 | g | ||
|
|
Sodium thioglycolate | 0.05 | g | ||
|
|
L-Cysteine HCl x H2O | 0.10 | g | ||
|
|
L-Asparagine | 0.25 | g | ||
|
|
Glucose | 0.20 | g | ||
|
|
Resazurin
(0.1%) |
0.10 | ml | ||
|
|
Distilled water | 94.00 | ml | ||
|
|
NaHCO3
(7.5%) |
2.67 | ml | ||
|
|
TPP/VFA mixture | 2.00 | ml | ||
|
|
Rabbit serum
(heat-inactivated at 56C for 1 hr) |
2.00 | ml | ||
TPP/VFA mixture #
11
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Thiamine pyrophosphate
(0.2%) |
1.5 | ml | ||
|
|
VFA solution | 1.0 | ml | ||
|
|
Distilled water | 8.4 | ml | ||
| 1 Mix together and filter-sterilize. Store at -20C. | |||||
VFA solution #
102
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Isobutyric acid | 0.5 | ml | ||
|
|
2-Methylbutyric acid | 0.5 | ml | ||
|
|
Isovaleric acid | 0.5 | ml | ||
|
|
Valeric acid | 0.5 | ml | ||
|
|
NaOH
(0.1 N) |
100.0 | ml | ||
| 1 Add volatile fatty acids to about 70-80 ml of NaOH solution and mix. Add NaOH solution to bring final volume to 100 ml. If not used the same day, filter-sterilize and store at 4C. | |||||