806 .
THERMOCOCCUS MEDIUM
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Equipment needed:
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Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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KCl | 0.33 | g | ||
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MgCl2 x 2 H2O | 2.70 | g | ||
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MgSO4 x 7 H2O | 3.40 | g | ||
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NH4Cl | 0.25 | g | ||
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CaCl2 x 2 H2O | 0.14 | g | ||
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K2HPO4 | 0.14 | g | ||
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Na2SeO3 | 1.00 | µg | ||
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NiCl2 x 6 H2O | 1.00 | µg | ||
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NaHCO3 | 1.00 | g | ||
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NaCl | 18.00 | g | ||
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Resazurin | 1.00 | mg | ||
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L-Cysteine HCl x H2O | 0.50 | g | ||
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Na2S x 9 H2O | 0.50 | g | ||
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Yeast extract | 1.00 | g | ||
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Trypticase | 1.00 | g | ||
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Sulphur | 5.00 | g | ||
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Modified Wolin's mineral solution II | 10.00 | ml | ||
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Wolin's vitamin solution | 10.00 | ml | ||
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Distilled water | 980.00 | ml | ||
| 1 Prepare the medium without NaHCO3, cysteine hydrochloride, Na2S x 9 H2O, and vitamin solution. Boil the medium and cool under N2. Add the NaHCO3 to the cooled medium and adjust the pH to about pH 7.2. Sterilise the medium at 100°C for 3 hours on 3 consecutive days. Reduce the medium by adding 10% neutralised sodium sulphide and cysteine hydrochloride from sterile stock solutions. The vitamins are added from a sterile (filter sterilised) stock solution prepared under nitrogen. The final pH is 7.2. | |||||
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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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. | |||||
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
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Biotin | 2.0 | mg |
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Folic acid | 2.0 | mg |
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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 |
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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 |