Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 1.00 | g | ||
|
KH2PO4 | 0.30 | g | ||
|
K2HPO4 | 0.30 | g | ||
|
Na2SO4 | 2.80 | g | ||
|
NaCl | 20.00 | g | ||
|
KCl | 0.10 | g | ||
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
Na2S2O3 x 5 H2O | 3.16 | g | ||
|
Yeast extract | 0.10 | g | ||
|
Trace element solution | 1.00 | ml | ||
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
Resazurin | 1.00 | mg | ||
|
Distilled water | 900.00 | ml | ||
1 To complete the medium, aseptically and anaerobically add the following stock solutions (per liter, final volume): | |||||
|
Sodium lactate
(1.0 M) |
20.00 | ml | ||
|
MgCl2 x 6 H2O
(15%) |
20.00 | ml | ||
|
NaHCO3
(5%) |
40.00 | ml | ||
|
Na2S x 9 H2O
(2%) |
20.00 | ml | ||
2 Mix ingredients and adjust pH to 7.4. Bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture tubes under the same gas mixture, seal with butyl rubber stoppers and autoclave. | |||||
3 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. | |||||
4 Readjust pH of the medium to 7.4, if necessary. |
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 |