860 .
ANAEROCOLUMNA MEDIUM
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* Medium for Clostridium thiosulfatireducens DSM 13105
Replace fructose with 2.5 g/l Na2S2O3 x 5 H2O and adjust pH of complete medium to 7.4.
Growth conditions: anaerobic
37 °C
Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
* 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.0 | g | ||
|
|
K2HPO4 | 0.3 | g | ||
|
|
KH2PO4 | 0.3 | g | ||
|
|
MgCl2 x 6 H2O | 0.2 | g | ||
|
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
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KCl | 0.1 | g | ||
|
|
NaCl | 0.6 | g | ||
|
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Yeast extract | 1.0 | g | ||
|
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Trace element solution SL-10 | 1.5 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
|
Na2CO3 | 1.5 | g | ||
| D-Fructose* | 4.0 | g | |||
|
|
L-Cysteine HCl x H2O | 0.5 | g | ||
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Na2S x 9 H2O | 0.3 | g | ||
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Na2S2O3 x 5 H2O | 2.5 | g | ||
|
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except carbonate, fructose, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fructose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.0 - 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 | ||
|
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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. | |||||