487 .
PEPTOCLOSTRIDIUM MEDIUM (BETAINE)
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Selected strain: Peptoclostridium litorale DSM 5388
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.3 - 7.5
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Betaine x H2O | 6.00 | g | ||
|
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L-Alanine | 2.20 | g | ||
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NaCl | 10.00 | g | ||
|
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MgSO4 x 7 H2O | 0.50 | g | ||
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NH4Cl | 1.00 | g | ||
|
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CaCl2 x 2 H2O | 0.03 | g | ||
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Trace element solution SL-10 | 1.00 | ml | ||
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Selenite-tungstate solution | 1.00 | ml | ||
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Yeast extract | 1.00 | g | ||
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Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
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NaHCO3 | 5.00 | g | ||
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K2HPO4 | 0.36 | g | ||
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KH2PO4 | 0.23 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Na2S x 9 H2O | 0.30 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except bicarbonate, phosphates, vitamins and sulfide), bring medium to the boil, then cool to room temperature under 100% N2 gas atmosphere. Add and dissolve bicarbonate, adjust pH to 7.5, dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add phosphates, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.3 - 7.5, if necessary. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
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FeCl2 x 4 H2O | 1.5 | g | ||
|
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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 | ||
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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. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
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Distilled water | 1000.0 | ml |
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
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 |
|
|
Distilled water | 1000 | ml |