1516 .
THERMOFILUM MEDIUM
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Equipment needed:
Hungate tubes
Final pH:
5.8 - 6.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Liquid medium | 1000.0 | ml | ||
| 1 Prepare the liquid medium. Boil the medium in a water bath and cool under N2 to room temperature. Adjust to pH 5.8-6.2. Dispense the medium into Hungate tubes or serum bottles under gassing with N2. Sterilize by autoclaving. Before inoculation add from sterile stock solutions: | |||||
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|
Filtrate of Desulfurococcus kamchatkensis (DSM18924) | 10.0 | ml | ||
|
|
Wolin's vitamin solution | 10.0 | ml | ||
|
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Na2S x H2O | 0.3 | g | ||
| 2 Prepare the liquid medium. Boil the medium in a water bath and cool under N2 to room temperature. Adjust to pH 5.8-6.2. Dispense the medium into Hungate tubes or serum bottles under gassing with N2. Sterilize by autoclaving. Before inoculation add from sterile stock solutions: | |||||
Trace Element Solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Fe(NH4)2(SO4)2 x 6 H2O | 784 | mg |
|
|
NiCl2 x 2 H2O | 225 | mg |
|
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CoCl2 x 6 H2O | 38 | mg |
|
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MnCl2 x 4 H2O | 99 | mg |
|
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ZnSO4 x 7 H2O | 144 | mg |
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Na2SeO4 | 95 | mg |
|
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Na2WO4 x 2 H2O | 33 | mg |
|
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Na2MoO4 x 2 H2O | 24 | mg |
|
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H3BO3 | 6 | mg |
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CuCl2 x 2 H2O | 2 | mg |
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Distilled water | 1000 | ml |
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
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 |
|
|
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 |
|
|
Distilled water | 1000.0 | ml |
Filtrate of Desulfurococcus kamchatkensis (DSM18924) #
10
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Liquid medium | 10.0 | ml | ||
|
|
Glucose | 2.0 | g/l | ||
|
|
Yeast extract | 0.6 | g/l | ||
| 1 Strain DSM 18924 was inoculated into anaerobic tube, containing 10ml of the medium mentioned above. Glucose (2g/l) and yeast extract (0,6g/l) were added as substrate. After 3 days of incubation culture was filtered throuth 0,22nm filter. Filtered culture broth is used for the cultivation of the strain at a concentration of 1% as a source of growth factor | |||||
Liquid medium #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 0.33 | g |
|
|
NH4Cl | 0.33 | g |
|
|
MgCl2 | 0.33 | g |
|
|
CaCl2 | 0.33 | g |
|
|
KCl | 0.33 | g |
|
|
Glucose | 2.00 | g |
|
|
Yeast extract | 0.10 | g |
|
|
Trace Element Solution | 1.00 | ml |
|
|
Distilled water | 1000.00 | ml |