842 .
HALOANAEROBACTER SALINARIUS MEDIUM
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Selected strain: Orenia salinaria DSM 13466
This strain uses the standard medium without modifications.
Growth conditions: 3-7 days
anaerobic
30 °C
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Ice bath
Final pH:
7.2 - 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 150.00 | g | ||
|
|
MgCl2 x 6 H2O | 15.00 | g | ||
|
|
KCl | 3.00 | g | ||
|
|
NH4Cl | 0.50 | g | ||
|
|
KH2PO4 | 0.35 | g | ||
|
|
CaCl2 x 2 H2O | 0.05 | g | ||
|
|
Yeast extract | 0.10 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
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NaHCO3 | 2.00 | g | ||
|
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Glucose | 1.20 | g | ||
|
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Resazurin | 1.00 | mg | ||
|
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Trace element solution SL-12 B | 1.00 | ml | ||
|
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Selenite-tungstate solution | 1.00 | ml | ||
|
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Vitamin solution 7 | 1.00 | ml | ||
|
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Distilled water | 997.00 | ml | ||
| 1 Final pH 7.2 - 7.4 | |||||
| 2 Prepare the medium without yeast extract, Na2S x 9 H2O, NaHCO3, glucose and vitamin solution (prepared as anaerobic, sterile stock solutions). Boil the medium and cool under anaerobic gas mixture of N2:CO2 (80:20). Dispense medium into Hungate tubes or serum bottles using the same gas mixture. Autoclave. Add the remaining components to the medium once it has cooled and check the final pH. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
NaOH | 0.4 | g |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution 7 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 10 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
Vitamin B12 | 50 | mg |
|
|
Thiamine | 100 | mg |
|
|
Distilled water | 1000 | ml |
Trace element solution SL-12 B #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 3.0 | g | ||
|
|
FeSO4 x 7 H2O | 1.1 | g | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
MnCl2 x 2 H2O | 50.0 | mg | ||
|
|
ZnCl2 | 42.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 18.0 | mg | ||
|
|
H3BO3 | 300.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Adjust pH to 6.0. | |||||