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DESULFATIBACILLUS OLEFINIVORANS MEDIUM
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Selected strain: Desulfatiferula olefinivorans DSM 18843
This strain uses the standard medium without modifications.
Equipment needed:
Autoclave
Filter
Final pH:
7.5
Select a strain (optional):
Solution A #
1005
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2SO4 | 3.00 | g | ||
|
|
KH2PO4 | 0.20 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
NaCl | 7.00 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
|
Yeast extract
(BD-Difco) |
0.10 | g | ||
|
|
Trace element solution SL12
(see below) |
1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
HEPES | 2.38 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 960.00 | ml | ||
| 1 Mix ingredients, adjust pH to 7.5 and autoclave under a N2 atmosphere. After cooling, add the following stock solutions (filter-sterilized or *autoclaved) to the solution: | |||||
|
|
MgCl2 x 6 H2O
(*, 15%) |
10.00 | ml | ||
|
|
NaHCO3
(8%) |
30.00 | ml | ||
|
|
Vitamin solution | 1.00 | ml | ||
|
|
Thiamine solution | 1.00 | ml | ||
|
|
Vitamin B12 solution 0.05 g/L | 1.00 | ml | ||
Substrate #
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Sodium palmitate | 2.8 | g | ||
| 1 Autoclave in culture vessels. | |||||
| 2 To complete the medium, combine Solution A and substrate in the culture vessels under a N2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Finally, reduce the medium with 0.01 volume of 5% Na2S x 9H2O solution (autoclaved and stored under a N2 atmosphere). | |||||
Trace metal solution SL12 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2-EDTA | 3.0 | g |
|
|
FeSO4 x 7 H2O | 1.1 | g |
|
|
CoCl2 x 6 H2O | 190.0 | mg |
|
|
MnCl2 x 4 H2O | 50.0 | mg |
|
|
ZnCl2 | 42.0 | mg |
|
|
NiCl2 x 6 H2O | 24.0 | mg |
|
|
Na2MoO4 x 2 H2O | 18.0 | mg |
|
|
H3BO3 | 300.0 | mg |
|
|
CuCl2 x 2 H2O | 2.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
p-Aminobenzoic acid | 4 | mg |
|
|
Biotin | 1 | mg |
|
|
Nicotinic acid | 10 | mg |
|
|
DL-Calcium pantothenate | 5 | mg |
|
|
Pyridoxine hydrochloride | 10 | mg |
|
|
Sodium phosphate buffer
(10 mM, pH 7.1) |
100 | ml |
Thiamine solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine HCl | 10 | mg |
|
|
Sodium phosphate buffer
(25 mM, pH 3.4) |
100 | ml |