291 .
PYROBACULUM NEUTROPHILUM MEDIUM
What is this page?
Equipment needed:
Filter
Gassing station
Serum bottle
Water bath
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 1.30 | g | ||
|
|
KH2PO4 | 0.28 | g | ||
|
|
MgSO4 x 7 H2O | 0.25 | g | ||
|
|
CaCl2 x 2 H2O | 0.07 | g | ||
|
|
FeCl3 x 6 H2O | 0.02 | g | ||
|
|
Allen's trace element solution | 10.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Prepare medium, then add per liter: | |||||
|
|
Sulfur
(powdered) |
8.00 | g | ||
|
|
Resazurin | 0.40 | mg | ||
| 2 Adjust pH to 6.5 with NaOH, bring to boil for 5 min, then cool to room temperature under H2 + CO2 (80% + 20%) gas mixture. Add 0.85 g NaHCO3 per liter and equilibrate the pH to c. 6.5 by further gassing. | |||||
| 3 Dispense in serum bottles (10 ml/30 ml bottle), seal and heat the bottles for one hour at 85°C on each of three successive days. | |||||
| 4 Before inoculation, add from anaerobic stock solutions 0.02% yeast extract (for mixotrophic growth only) and 0.01 ml Na-dithionite solution (250 mg Na-dithionite/10 ml dist. water, filter-sterilized). After inoculation, pressurize to 1 bar H2 + CO2 overpressure. | |||||
Allen's trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MnCl2 x 4 H2O | 180 | mg | ||
|
|
Na2B4O7 x 10 H2O | 450 | mg | ||
|
|
ZnSO4 x 7 H2O | 22 | mg | ||
|
|
CuCl2 x 2 H2O | 5 | mg | ||
|
|
Na2MoO4 x 2 H2O | 3 | mg | ||
|
|
VOSO4 x 2 H2O | 3 | mg | ||
|
|
CoSO4 x 7 H2O | 1 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH of final solution to 2 with 1 N HCl. | |||||