461 .
MINERAL MEDIUM (NAGEL AND ANDREESEN)
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Equipment needed:
Autoclave
Filter
Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Distilled water | 1000.00 | ml | ||
|
|
Na2HPO4 x 2 H2O | 1.45 | g | ||
|
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KH2PO4 | 0.25 | g | ||
|
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CaCl2 | 0.01 | g | ||
|
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MgSO4 x 7 H2O | 0.50 | g | ||
|
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MnSO4 | 0.01 | g | ||
|
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NH4Cl | 0.30 | g | ||
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NaCl | 0.05 | g | ||
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Vitamin solution | 5.00 | ml | ||
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Trace element solution SL-10 | 1.00 | ml | ||
| 1 Dissolve and autoclave the phosphates separately. Add the vitamin solution and trace element solution after autoclaving. After mixing, pH should be 7.5. | |||||
| 2 Rehydrate and cultivate the lyophilized cells in complex medium recommended for the specific strain (e.g. medium 1, 220 or 535) first. After this reactivation, cultivate on mineral medium 461 with the appropriate carbon source. | |||||
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Vitamin B12 | 50 | mg | ||
|
|
Pantothenic acid | 50 | mg | ||
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Riboflavin | 50 | mg | ||
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Pyridoxamine hydrochloride | 10 | mg | ||
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Biotin | 20 | mg | ||
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Folic acid | 20 | mg | ||
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Nicotinic acid | 25 | mg | ||
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Nicotine amide | 25 | mg | ||
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alpha-lipoic acid | 50 | mg | ||
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p-Aminobenzoic acid | 50 | mg | ||
|
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Thiamine-HCl x 2 H2O | 50 | mg | ||
|
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Distilled water | 1000 | ml | ||
| 1 Stir for some hours, filter sterilize the solution. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
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MnCl2 x 4 H2O | 100.0 | mg | ||
|
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H3BO3 | 6.0 | mg | ||
|
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CoCl2 x 6 H2O | 190.0 | mg | ||
|
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CuCl2 x 2 H2O | 2.0 | mg | ||
|
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NiCl2 x 6 H2O | 24.0 | mg | ||
|
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||