1098 .
ARCHAEOGLOBUS INFECTUS MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Sea Salt
(SIGMA) |
35.00 | g | ||
|
|
NH4Cl | 0.10 | g | ||
|
|
KH2PO4 | 0.05 | g | ||
|
|
Na-acetate | 1.60 | g | ||
|
|
Yeast extract | 0.10 | g | ||
|
|
Wolfe's mineral elixir | 1.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
NaHCO3 | 2.00 | g | ||
|
|
2-Mercaptoethanesulfonic acid
(coenzyme M) |
0.14 | g | ||
|
|
Seven vitamins solution | 1.00 | ml | ||
|
|
Na2S2O3 x 5 H2O | 2.50 | g | ||
|
|
DL-Dithiothreitol | 0.30 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, thiosulfate, 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., Balch-type tubes) to 30% of their volume and autoclave. Add coenzyme M, vitamins, thiosulfate, and dithiothreitol 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 atmosphere. The vitamin and dithiothreitol solutions should be sterilized by filtration. Adjust the pH of the complete medium to 6.5. | |||||
| 2 After inoculation pressurize culture vessels to 1.5 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
Wolfe's mineral elixir #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MgSO4 x 7 H2O | 30.00 | g | ||
|
|
MnSO4 x H2O | 5.00 | g | ||
|
|
NaCl | 10.00 | g | ||
|
|
FeSO4 x 7 H2O | 1.00 | g | ||
|
|
CoCl2 x 6 H2O | 1.80 | g | ||
|
|
CaCl2 x 2 H2O | 1.00 | g | ||
|
|
ZnSO4 x 7 H2O | 1.80 | g | ||
|
|
CuSO4 x 5 H2O | 0.10 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.18 | g | ||
|
|
H3BO3 | 0.10 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.10 | g | ||
|
|
(NH4)2Ni(SO4)2 x 6 H2O | 2.80 | g | ||
|
|
Na2WO4 x 2 H2O | 0.10 | g | ||
|
|
Na2SeO4 | 0.10 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts. | |||||
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
|
D-(+)-biotin | 20 | mg |
|
|
Nicotinic acid | 200 | mg |
|
|
Calcium pantothenate | 100 | mg |
|
|
Pyridoxine hydrochloride | 300 | mg |
|
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
|
Distilled water | 1000 | ml |