716 .
AMMONIFEX DEGENSII MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Serum bottle
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K2HPO4 x 3 H2O | 0.30 | g | ||
|
|
KH2PO4 | 0.22 | g | ||
|
|
(NH4)2SO4 | 0.22 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
MgSO4 x 7 H2O | 0.09 | g | ||
|
|
CaCl2 x 2 H2O | 0.06 | g | ||
|
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
2.00 | ml | ||
|
|
NiCl2 x 6 H2O
(0.1% w/v) |
0.20 | ml | ||
|
|
Modified Wolin's mineral solution | 1.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
KNO3 | 1.00 | g | ||
|
|
Na2CO3 | 1.50 | g | ||
|
|
DL-Dithiothreitol | 0.30 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except nitrate, carbonate, and dithiothreitol), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic serum vials (e.g., 10 ml medium in 25 ml Balch-type tubes) and autoclave. Before use, add nitrate from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Before inoculation add dithiothreitol from an anoxic stock solution sterilized by filtration under 100% N2 gas and adjust the pH of the medium to 7.2. | |||||
| 2 After growth has started pressurize vials to 0.5 bar overpressure with a sterile 80% H2 and 20% CO2 gas mixture. | |||||
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||