492 .
CLOSTRIDIUM METHYLPENTOSUM MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
6.8 - 7.0
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Solution A | 954 | ml | ||
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Solution B | 20 | ml | ||
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Solution C | 10 | ml | ||
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Solution D | 1 | ml | ||
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Solution E | 20 | ml | ||
| 1 Sparge solution A with 100% N2 gas for at least 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution E is autoclaved separately under 100% N2 gas. Solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 6.8 - 7.0. | |||||
Solution A #
954
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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NH4Cl | 0.9 | g | ||
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KH2PO4 | 0.9 | g | ||
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NaCl | 0.9 | g | ||
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CaCl2 x 2 H2O | 20.0 | mg | ||
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MnCl2 x 4 H2O | 20.0 | mg | ||
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MgSO4 x 7 H2O | 20.0 | mg | ||
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CoCl2 x 6 H2O
(0.1% w/v) |
5.0 | ml | ||
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FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
5.0 | ml | ||
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ZnSO4 x 7 H2O
(0.1% w/v) |
2.0 | ml | ||
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CuSO4 x H2O
(0.1% w/v) |
2.0 | ml | ||
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Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
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Distilled water | 940.0 | ml | ||
| 1 Adjust pH to 6.5 with KOH. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
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Folic acid | 20 | mg |
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Pyridoxine hydrochloride | 100 | mg |
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Thiamine HCl | 50 | mg |
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Riboflavin | 50 | mg |
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Nicotinic acid | 50 | mg |
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Calcium D-(+)-pantothenate | 50 | mg |
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Vitamin B12 | 1 | mg |
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p-Aminobenzoic acid | 50 | mg |
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(DL)-alpha-Lipoic acid | 50 | mg |
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Distilled water | 1000 | ml |