565 .
DESULFOVIBRIO HEB233 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
7.2
Select a strain (optional):
Solution A #
902
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.3 | g | ||
|
|
K2HPO4 | 0.3 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
Na2SO4 | 3.0 | g | ||
|
|
NaCl | 23.0 | g | ||
|
|
KCl | 0.1 | g | ||
|
|
CaCl2 | 0.1 | g | ||
|
|
Yeast extract
(BD-Difco) |
0.1 | g | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
|
Resazurin | 1.0 | mg | ||
|
|
Distilled water | 900.0 | ml | ||
| 1 To complete the medium, add the following stock solutions [Filter-sterilize NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. The other solutions are autoclaved and stored anaerobically under a N2 atmosphere] to the medium (per 4.5 ml): | |||||
|
|
Sodium lactate
(1 M) |
0.1 | ml | ||
|
|
MgCl2 x 6 H2O
(15%) |
0.1 | ml | ||
|
|
NaHCO3
(5%) |
0.2 | ml | ||
|
|
Na2S x 9 H2O
(2%) |
0.1 | ml | ||
| 2 Mix ingredients and adjust pH to 7.2. Distribute 0.9 volume of the medium into tubes (e.g., 4.5 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave at 110C for 45 min. | |||||
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 |