439 .
DESULFOVIBRIO AEROTOLERANS DVO5 MEDIUM
What is this page?
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.00 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KH2PO4 | 2.00 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
Yeast extract | 0.50 | g | ||
|
|
Na2SO4 | 4.00 | g | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components thoroughly and adjust pH to 7.0. Bring to a boil, and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 10 ml medium in Hungate tubes) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, anaerobically and aseptically add per liter the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
Sodium lactate
(1.0 M) |
20.00 | ml | ||
|
|
Vitamin solution
(*) |
1.00 | ml | ||
|
|
Thiamine solution
(*) |
1.00 | ml | ||
|
|
Vitamin B12 solution 0.05 g/L
(*) |
1.00 | ml | ||
|
|
NaHCO3
(*, 8.0%) |
20.00 | ml | ||
| 2 Prior to use, add per liter the following solution (autoclaved and stored under a N2 atmosphere): | |||||
|
|
Na2S x 9 H2O
(5%) |
2.00 | ml | ||
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(25%) |
12.5 | ml |
|
|
FeSO4 x 7 H2O | 2.1 | g |
|
|
H3BO3 | 30.0 | mg |
|
|
MnCl2 x 4 H2O | 100.0 | mg |
|
|
CoCl2 x 6 H2O | 190.0 | mg |
|
|
NiCl2 x 6 H2O | 24.0 | mg |
|
|
CuCl2 x 2 H2O | 2.0 | mg |
|
|
ZnSO4 x 7 H2O | 144.0 | mg |
|
|
Na2MoO4 x 2 H2O | 36.0 | mg |
|
|
Distilled water | 987.0 | 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 |