705 .
BACILLUS INFERNUS MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Ferric citrate monohydrate | 5.3 | g | ||
|
|
NaCl | 5.8 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
K2HPO4 x 3 H2O | 0.4 | g | ||
|
|
MgCl2 x 6 H2O | 1.0 | g | ||
|
|
Yeast extract
(OXOID) |
2.0 | g | ||
|
|
Trypticase peptone
(BD BBL) |
2.0 | g | ||
|
|
Trace element solution | 10.0 | ml | ||
|
|
Na-acetate | 0.8 | g | ||
|
|
Na-lactate | 1.2 | g | ||
|
|
2-Mercaptoethanesulfonic acid
(coenzyme M) |
0.5 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 First, dissolve ferric citrate in water by heating and adjust to pH 7.5 with NaOH, then add other medium ingredients except acetate, lactate, and coenzyme M. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add acetate, lactate, and coenzyme M from sterile anoxic stock solutions prepared under 100% N2 gas. | |||||
| 2 Prior to inoculation the pH of the complete medium should be checked and adjusted to 7.5, if necessary. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 0.50 | g | ||
|
|
CoCl2 x 6 H2O | 0.15 | g | ||
|
|
MnCl2 x 4 H2O | 0.10 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
ZnCl2 | 0.10 | g | ||
|
|
AlCl3 x 6 H2O | 40.00 | mg | ||
|
|
Na2WO4 x 2 H2O | 40.00 | mg | ||
|
|
Na2SeO3 x 5 H2O | 30.00 | mg | ||
|
|
NiCl2 x 6 H2O | 20.00 | mg | ||
|
|
CuCl2 x 2 H2O | 20.00 | mg | ||
|
|
H3BO3 | 10.00 | mg | ||
|
|
Na2MoO4 x 2 H2O | 10.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7 using 2 N NaOH, then dissolve remaining compounds. | |||||