480b .
TTD-MEDIUM
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Selected strain: Thermococcus stetteri DSM 5262
This strain uses the standard medium without modifications.
Growth conditions: 75 °C
Equipment needed:
Filter
Hungate tubes
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.33 | g | ||
|
|
CaCl2 x 2 H2O | 0.33 | g | ||
|
|
MgCl2 x 6 H2O | 0.33 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
KH2PO4 | 0.33 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Sulfur
(powder) |
10.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Add to the medium 0.5% casitone and 2.5% NaCl (marine source). Adjust pH to 5.7 with hydrochloric acid before sterilization. Sterilize the medium by heating for 1 h at 90 - 100°C on three successive days. Before use add 30.0 ml/l of 5% (w/v) sterile, anaerobic NaHCO3-solution, vitamins and sulfide. Final pH should be 6.5. | |||||
|
|
Casitone | 0.50 | % | ||
|
|
NaCl | 2.50 | % | ||
|
|
NaHCO3
(5% w/v) |
30.00 | ml | ||
|
|
Wolin's vitamin solution | 10.00 | ml | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
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 |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |