81a .
THIOMONAS ISLANDICA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Final pH:
5.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K2HPO4 | 2.30 | g | ||
|
|
Na2HPO4 x 2 H2O | 2.90 | g | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
CaCl2 x 2 H2O | 0.01 | g | ||
|
|
MgSO4 x 7 H2O | 0.50 | g | ||
|
|
NaVO3 x H2O
(0.1% w/v) |
5.00 | ml | ||
|
|
Trace element solution SL-6 | 5.00 | ml | ||
|
|
NaHCO3 | 0.50 | g | ||
|
|
Ferric ammonium citrate | 0.05 | g | ||
|
|
Seven vitamins solution | 1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (ecxept bicarbonate, ferric ammonium citrate and vitamins), then sparge solution with 80% N2 and 20% CO2 gas mixture for at least 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO2. Adjust pH to 4.5 and dispense solution under air atmosphere into serum vials suitable for anaerobic cultivation to 50% of volume. Close vials with butyl rubber septa to prevent free exchange of oxygen with the external atmosphere and autoclave. Add ferric ammonium citrate and vitamins from anoxic stock solutions sterilized by filtration and bicarbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Adjust pH of the complete medium to 5.0, if necessary. | |||||
| 2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure. | |||||
| 3 Note: Use at least 10% (v/v) as inoculum. | |||||
Trace element solution SL-6 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnSO4 x 7 H2O | 0.10 | g |
|
|
MnCl2 x 4 H2O | 0.03 | g |
|
|
H3BO3 | 0.30 | g |
|
|
CoCl2 x 6 H2O | 0.20 | g |
|
|
CuCl2 x 2 H2O | 0.01 | g |
|
|
NiCl2 x 6 H2O | 0.02 | g |
|
|
Na2MoO4 x 2 H2O | 0.03 | g |
|
|
Distilled water | 1000.00 | ml |
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
|
D-(+)-biotin | 20 | mg |
|
|
Nicotinic acid | 200 | mg |
|
|
Calcium pantothenate | 100 | mg |
|
|
Pyridoxine hydrochloride | 300 | mg |
|
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
|
Distilled water | 1000 | ml |