701 .
HALOANAEROBACTER CHITINOVORANS MEDIUM
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Equipment needed:
Autoclave
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.0 | g | ||
|
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MgCl2 x 6 H2O | 7.0 | g | ||
|
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MgSO4 x 7 H2O | 9.6 | g | ||
|
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CaCl2 x 2 H2O | 0.5 | g | ||
|
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KCl | 3.8 | g | ||
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K2HPO4 x 3 H2O | 0.4 | g | ||
|
|
NaCl | 10.0 | g | ||
|
|
NaHCO3 | 3.0 | g | ||
|
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Na2CO3 | 1.0 | g | ||
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Na2S x 9 H2O | 0.5 | g | ||
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Yeast extract | 1.0 | g | ||
|
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L-Cysteine | 0.5 | g | ||
|
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Resazurin | 1.0 | mg | ||
|
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Substrate
(glucose, chitin etc.) |
5.0 | g | ||
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Modified Wolin's mineral solution II | 1.0 | ml | ||
|
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Distilled water | 1000.0 | ml | ||
| 1 pH 7.2 | |||||
| 2 Prepare the medium without the Na2S, Na2CO3, NaHCO3, L-cysteine and substrate. Boil the medium and cool under a stream of nitrogen. Add the Na2CO3 and NaHCO3 . Adjust the pH to 7.2. If chitin is used as the substrate it must be added before autoclaving. Autoclave the medium and add the Na2S (from a sterile, neutralized stock solution prepared under nitrogen), L-cysteine (from a stock solution prepared under nitrogen), and the glucose or N-acetylglucosamine (if used as substrate) from sterile stock solution prepared under nitrogen. Check the final pH. | |||||
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.500 | g | ||
|
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MgSO4 x 7 H2O | 3.000 | g | ||
|
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MnSO4 x H2O | 0.500 | g | ||
|
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NaCl | 1.000 | g | ||
|
|
FeSO4 x 7 H2O | 0.100 | g | ||
|
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CoSO4 x 7 H2O | 0.180 | g | ||
|
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CaCl2 x 2 H2O | 0.100 | g | ||
|
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ZnSO4 x 7 H2O | 0.180 | g | ||
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CuSO4 x 5 H2O | 0.010 | g | ||
|
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AlK(SO4)2 x 12 H2O | 0.020 | g | ||
|
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H3BO3 | 0.010 | g | ||
|
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Na2MoO4 x 2 H2O | 0.010 | g | ||
|
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NiCl2 x 6 H2O | 0.025 | g | ||
|
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Na2SeO3 x 5 H2O | 0.300 | mg | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||