926 .
ALKALIPHILIC THERMOCOCCUS MEDIUM
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* Medium for Thermococcus acidaminovorans DSM 11906
To the sterile, anaerobic, mineral medium add 0.08 ml polysulphide/10 ml medium, casamino acids to a final concentration of 0.2%. The may be precipitation of material and the medium will turn pale yellow due to the addition of the polysulphide. The colour will disappear as the strain grows.
Growth conditions: 12-24 h
anaerobic
85 °C
pH 7.50
Equipment needed:
Anaerobic workbench
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 27.70 | g | ||
|
|
MgSO4 x 7 H2O | 7.00 | g | ||
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MgCl2 x 6 H2O | 5.50 | g | ||
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KCl | 0.65 | g | ||
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NaBr | 0.10 | g | ||
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NaHCO3 | 0.32 | g | ||
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K2HPO4 | 1.00 | g | ||
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CaCl2 x 2 H2O | 0.05 | mg | ||
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KI | 15.00 | mg | ||
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H3BO3 | 0.03 | g | ||
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Modified Wolin's mineral solution II | 20.00 | ml | ||
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Casamino acids | 0.20 | % | ||
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Distilled water | 2000.00 | ml | ||
| 1 Prepare the medium anaerobically under nitrogen. Do not adjust the pH. | |||||
| 2 Prepare separate anaerobic stock solutions of, Casamino acids(10%), Yeast extract (10%), and glycine (2M = 150 g/l). A 0.5M polysulphide solution is prepared by dissolving 12.0 g Na2S x 9 H2O in oxygen free water, followed by adding 1.6 g sulphur - the solution will be dark yellow. | |||||
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 | ||
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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 | ||
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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. | |||||