1600 .
FRANCISELLA PERSICA MEDIUM
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* Medium for Francisella persica DSM 101678
Plus 5% CO2
Growth conditions: 3-7 days
34 °C
Equipment needed:
Filter
Final gas composition:
5 % CO2
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Final pH:
6.8
Select a strain (optional):
ml
| 1 2x concentrated stock: | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
L-15 powder
(Leibovitz medium) |
1.000 | pack | ||
|
|
DL-aspartic acid | 0.299 | g | ||
|
|
L-Glutamic acid | 0.500 | g | ||
|
|
L-Glutamine
(200 mM) |
10.000 | ml | ||
|
|
L-Proline | 0.300 | g | ||
|
|
alpha-ketoglutaric acid | 0.299 | g | ||
|
|
D-Glucose | 2.239 | g | ||
|
|
Stock Solution D (10 ml) | 1.000 | ml | ||
|
|
Vitamin Stock (10 ml) | 1.000 | ml | ||
|
|
Demineralized water | 346.000 | ml | ||
| 2 Adjust pH to 6.8 with NaOH. | |||||
| 3 When ready to use add: | |||||
| 4 Aliquot 17.3ml into 50ml tubes and store at -20°C. | |||||
| 5 Fill up to 346ml with demineralized water and filter sterilize through 0.2µm filter | |||||
|
|
Demineralized water
(sterile ) |
25.000 | ml | ||
|
|
Tryptose-phosphate
(broth) |
5.000 | ml | ||
|
|
FBS
(heat inactivated) |
2.500 | ml | ||
|
|
cholesterol lipid concentrate
(250x, ThermoFisher Scientific) |
200.000 | µl | ||
| 6 Filter sterilize each tube through 0.2µm filter. | |||||
Stock Solution A (10 ml) #
10
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnSO4 x 7 H2O | 0.20 | g |
|
|
MnSO4 x 1 H2O | 0.16 | g |
|
|
CuSO4 x 5 H2O | 0.02 | g |
|
|
CoCl2 x 6 H2O | 0.02 | g |
Stock Solution D (10 ml) #
10
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Glutathione
(reduced) |
1.00 | g |
|
|
Ascorbic acid | 1.00 | g |
|
|
FeSO4 x 7 H2O | 0.05 | g |
|
|
Stock Solution A (10 ml) | 0.10 | ml |
|
|
Stock Solution B (10 ml) | 0.10 | ml |
|
|
Stock Solution C (10 ml) | 0.10 | ml |
Vitamin Stock (10 ml) #
10
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Para-aminobenzoic acid | 0.10 | g |
|
|
Vitamin B12 | 0.05 | g |
|
|
D-Biotin | 0.01 | g |