Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 1.0 | g | ||
|
K2HPO4 | 0.5 | g | ||
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
KCl | 0.8 | g | ||
|
Sodium acetate | 0.5 | g | ||
|
NaCl | 200.0 | g | ||
|
Yeast extract
(BD-Difco) |
1.0 | g | ||
|
Tryptone
(BD-Difco) |
0.5 | g | ||
|
Trace element solution | 1.0 | ml | ||
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
Resazurin | 0.5 | mg | ||
|
Distilled water | 855.0 | ml | ||
1 Mix components thoroughly, adjust pH to 7.6 with KOH and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions from anaerobic stocks (autoclaved or *filter-sterilized): | |||||
|
NaHCO3
(*, 8%) |
25.0 | ml | ||
|
Glucose
(1 M) |
20.0 | ml | ||
|
MgSO4 x 7 H2O
(10%) |
50.0 | ml | ||
|
Tryptone
(10%) |
50.0 | ml | ||
2 Aseptically distribute the medium into culture vessels under a stream of N2-CO2 (4:1, v/v), and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution from an anaerobic stock (autoclave and store under a N2 atmosphere). | |||||
|
Na2S x 9 H2O
(5%) |
8.0 | ml |
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 |