591 .
HALOBACTEROIDES/HALOINCOLA MEDIUM
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* Medium for Halanaerobacter lacunarum DSM 6640
Use 150 g/l NaCl, replace the peptone with 0.5 g/l yeast extract (Difco) and adjust the pH to 7.5.
Growth conditions: 37 °C
Equipment needed:
Autoclave
Filter
Final pH:
* 7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.33 | g | ||
|
|
CaCl2 | 0.33 | g | ||
|
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MgCl2 | 0.33 | g | ||
|
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KCl | 0.33 | g | ||
|
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KH2PO4 | 0.33 | g | ||
|
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NaCl , adjust if required in range 100-150 | 150.00 | g/l | ||
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NaHCO3 | 1.50 | g | ||
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Na2S x 9 H2O | 0.50 | g | ||
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Glucose | 5.00 | g | ||
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Peptone*
(Bacto Proteose Peptone No.3) |
5.00 | g | |||
|
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Trace element solution SL-10
(SL 10) |
1.00 | ml | ||
|
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Wolin's vitamin solution | 10.00 | ml | ||
|
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Resazurin | 2.00 | mg | ||
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Yeast extract | 0.50 | g/l | ||
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Distilled water | 990.00 | ml | ||
| 1 Prepare the medium without NaHCO3, Na2S, glucose and vitamins, boil and cool under a nitrogen:carbon dioxide (80:20 v/v) atmosphere. Glucose (filter-sterilized and stored under nitrogen), vitamins (filter sterilized and stored under nitrogen), and sodium sulphide (autoclaved under nitrogen) are added from sterile stock solutions. The pH should be pH 7.5. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
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 | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
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. | |||||
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
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Thiamine HCl | 5.0 | mg |
|
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Riboflavin | 5.0 | mg |
|
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Nicotinic acid | 5.0 | mg |
|
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
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Vitamin B12 | 0.1 | mg |
|
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p-Aminobenzoic acid | 5.0 | mg |
|
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(DL)-alpha-Lipoic acid | 5.0 | mg |
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Distilled water | 1000.0 | ml |