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CALDICELLULOSIRUPTOR MEDIUM
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Selected strain: Thermoanaerobacter ethanolicus DSM 2246
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
65 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.2
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.90 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
KH2PO4 | 0.75 | g | ||
|
|
K2HPO4 | 1.50 | g | ||
|
|
Trypticase peptone
(BD BBL) |
2.00 | g | ||
|
|
Yeast extract
(OXOID) |
1.00 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
FeCl3 x 6 H2O
(0.1% w/v in 0.2 N HCl) |
2.50 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.75 | g | ||
|
|
Cellobiose | 1.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except cysteine and cellobiose. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then add cysteine and adjust pH to 7.2. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose after autoclaving from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of the complete medium to 7.2, if necessary. | |||||
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 | ||
|
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
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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. | |||||