428 .
HETEROTROPHIC MEDIUM H3P
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Equipment needed:
Autoclave
Filter
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Solution A | 50 | ml | ||
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Solution B | 855 | ml | ||
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Solution C | 20 | ml | ||
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Solution D | 30 | ml | ||
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Solution E | 50 | ml | ||
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Standard vitamin solution | 5 | ml | ||
| 1 H3P is a heterotrophic medium for growth, purity checking and isolation of a broad spectrum of aerobic bacteria (Ref. 3367). | |||||
| 2 Solutions A, B, C and D are autoclaved separately for 15 min at 121°C, cooled down to 50°C and then mixed aseptically with filter-sterilized solution E (at 50°C) and 5.0 ml of standard vitamin solution. The final pH of this medium should be 6.8 without adjustment. | |||||
Solution B #
855
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NH4Cl | 1.00 | g |
|
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MgSO4 x 7 H2O | 0.50 | g |
|
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CaCl2 x 2 H2O | 0.01 | g |
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MnCl2 x 4 H2O | 5.00 | mg |
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NaVO3 x H2O | 5.00 | mg |
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Trace element solution SL-6 | 5.00 | ml |
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Distilled water | 850.00 | ml |
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Agar , if required | 20.00 | g |
Solution D #
30
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Yeast extract | 1 | g | ||
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Na-acetate | 1 | g | ||
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Disodium succinate | 1 | g | ||
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DL-Malate | 1 | g | ||
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Distilled water | 30 | ml | ||
| 1 pH 7.0 | |||||
Solution E #
50
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Na-lactate | 1 | g | ||
|
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Na-pyruvate | 1 | g | ||
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D-Mannitol | 1 | g | ||
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D-Glucose | 2 | g | ||
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Distilled water | 50 | ml | ||
| 1 pH 7.0 | |||||
| 2 filter-sterilized | |||||
Standard vitamin solution #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Riboflavin | 10.0 | mg | ||
|
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Thiamine-HCl x 2 H2O | 50.0 | mg | ||
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Nicotinic acid | 50.0 | mg | ||
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Pyridoxine hydrochloride | 50.0 | mg | ||
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Calcium pantothenate | 50.0 | mg | ||
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Biotin | 0.1 | mg | ||
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Folic acid | 0.2 | mg | ||
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Vitamin B12 | 1.0 | mg | ||
|
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Distilled water | 100.0 | ml | ||
| 1 Filter sterilize (0.2 µm pore-size) and store at 4°C. | |||||
Trace element solution SL-6 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnSO4 x 7 H2O | 0.10 | g |
|
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MnCl2 x 4 H2O | 0.03 | g |
|
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H3BO3 | 0.30 | g |
|
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CoCl2 x 6 H2O | 0.20 | g |
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CuCl2 x 2 H2O | 0.01 | g |
|
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NiCl2 x 6 H2O | 0.02 | g |
|
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Na2MoO4 x 2 H2O | 0.03 | g |
|
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Distilled water | 1000.00 | ml |