358a .
ACIDIANUS MEDIUM (ANAEROBIC)
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* Medium for Stygiolobus azoricus DSM 6296
Reduce amount of yeast extract to 0.20 g/l and adjust pH to 2.5 - 3.0. Pressurize inoculated bottles to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Growth conditions: anaerobic
80 °C
Equipment needed:
Autoclave
Gassing station
Serum bottle
Water bath
Final pH:
* 2.5 - 3.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 1.30 | g | ||
|
|
KH2PO4 | 0.28 | g | ||
|
|
MgSO4 x 7 H2O | 0.25 | g | ||
|
|
CaCl2 x 2 H2O | 0.07 | g | ||
|
|
FeCl3 x 6 H2O | 0.02 | g | ||
|
|
Allen's trace element solution | 10.00 | ml | ||
|
|
Sulfur
(powder) |
5.00 | g | ||
|
|
Yeast extract
(OXOID) |
0.20 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients, except sulfur and yeast extract, bring medium to the boil, then cool to room temperature under 80% H2 and 20% CO2 gas mixture and adjust pH to 2.5 using 10 N H2SO4. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml serum bottles) containing already the appropriate amount of sulfur. For sterilization sealed bottles with medium are heated in a boiling water bath for 2 - 3 h on each of 3 successive days. Add yeast extract from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. | |||||
| 2 Pressurize inoculated bottles to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
| 3 Note: Inoculate with 5% (w/v) culture. Incubate without shaking. | |||||
Allen's trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MnCl2 x 4 H2O | 180 | mg | ||
|
|
Na2B4O7 x 10 H2O | 450 | mg | ||
|
|
ZnSO4 x 7 H2O | 22 | mg | ||
|
|
CuCl2 x 2 H2O | 5 | mg | ||
|
|
Na2MoO4 x 2 H2O | 3 | mg | ||
|
|
VOSO4 x 2 H2O | 3 | mg | ||
|
|
CoSO4 x 7 H2O | 1 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH of final solution to 2 with 1 N HCl. | |||||