929 .
RHODOVULUM IODOSUM AND R. ROBIGINOSUM MEDIUM
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Equipment needed:
Anaerobic workbench
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 26.40 | g | ||
|
|
MgCl2 x 6 H2O | 5.70 | g | ||
|
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MgSO4 x 7 H2O | 6.80 | g | ||
|
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CaCl2 x 2 H2O | 1.50 | g | ||
|
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KCl | 0.66 | g | ||
|
|
KBr | 0.09 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Prepare the medium anaerobically under an atmosphere of N2:CO2 (8:2 vol:vol) and then add: | |||||
|
|
NaHCO3
(84 g/l autoclaved under a CO2 atmopshere) |
30.00 | ml/l | ||
|
|
KH2PO4 | 0.40 | g/l | ||
|
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NH4Cl | 0.25 | g/l | ||
|
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Na2S2O3 | 0.50 | mM | ||
|
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Trace elements | 1.00 | ml/l | ||
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Selenite/tungsate solution | 1.00 | ml/l | ||
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Vitamin solution | 1.00 | ml/l | ||
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Vitamin B12 solution 0.05 g/L | 1.00 | ml/l | ||
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Vitamin B1 solution 0.1 g/L | 1.00 | ml/l | ||
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Acetate | 5.00 | mM | ||
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FeSO4 | 10.00 | mM | ||
| 2 The pH is adjusted to 6.8 | |||||
| 3 To the medium add 5 mM acetate and 10 mM FeSO4 (from a 1 M stock solution kept under nitrogen). When the iron is added to the medium a white floculant precipitate may appear. In the initial stages of growth the precipitate turns orange/brown as the ferrous ions are oxidised to ferrihydrite. Cultivation may take 2-3 weeks at a distance of 30 cm from a 25 W light bulb. | |||||
Trace elements #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
FeSO4 x 7 H2O | 2.1 | g | ||
|
|
Na2-EDTA | 5.2 | g | ||
|
|
H3BO3 | 30.0 | mg | ||
|
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MnCl2 x 4 H2O | 100.0 | mg | ||
|
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CoCl2 x 6 H2O | 190.0 | mg | ||
|
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NiCl2 x 6 H2O | 24.0 | mg | ||
|
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CuCl2 x 2 H2O | 2.0 | mg | ||
|
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ZnSO4 x 7 H2O | 144.0 | mg | ||
|
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
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Distilled water | 1000.0 | ml | ||
| 1 Adjust the pH to 6.0 | |||||
Selenite/tungsate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2SeO3 x 5 H2O | 6 | mg |
|
|
Na2WO4 x 2 H2O | 8 | mg |
|
|
NaOH | 200 | mg |
|
|
Distilled water | 1000 | ml |
Vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Para-aminobenzoic acid | 4 | mg |
|
|
Biotin | 1 | mg |
|
|
Nicotinate | 10 | mg |
|
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Pantothenate | 5 | mg |
|
|
Pyridoxin | 15 | mg |
|
|
Distilled water | 100 | ml |
Vitamin B12 solution 0.05 g/L #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Cyanocobalamine | 5 | mg |
|
|
Distilled water | 100 | ml |