470 .
VULCANIBACILLUS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
1.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Sea Salt
(Sigma) |
30.00 | g | ||
|
|
NaNO3 | 1.00 | g | ||
|
|
Wolfe's mineral elixir | 2.00 | ml | ||
|
|
VOSO4 x n H2O | 0.05 | mg | ||
|
|
Sodium pyruvate | 1.00 | g | ||
|
|
Resazurin | 0.50 | mg | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
NaHCO3 | 1.00 | g | ||
|
|
Yeast extract | 0.50 | g | ||
|
|
Trace vitamins | 20.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except FeSO4 x 7H2O, NaHCO3, yeast extract and vitamins, and adjust pH to 7.0. Bring to a boil for 3 min, then cool to room temperature under a N2 gas stream. Dispense the medium under the same gas atmosphere into culture vessels, seal with butyl rubber stoppers and autoclave. Add sterile anaerobic stock solutions of FeSO4 x 7H2O (1%, in 0.1 N H2SO4), NaHCO3 (8%, under a N2 x CO2 gas mixture), yeast extract (10%) and vitamins (filter-sterilized) to the medium. Adjust final pH of the medium to 7.0 - 7.2. Add 10 - 20 mg sodium dithionite per liter [e.g., from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized], if the medium is not completely reduced after inoculation. | |||||
Wolfe's mineral elixir #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MgSO4 x 7 H2O | 30.00 | g | ||
|
|
MnSO4 x n 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 | ||
|
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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 dissolve the salts. | |||||
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |