Microaerophilic with 5% oxygen
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 120.0 | g | ||
|
K2HPO4 | 2.0 | g | ||
|
(NH4)2SO4 | 0.5 | g | ||
|
Distilled water | 1000.0 | ml | ||
1 Flush the gas phase with nitrogen and add 5% oxygen. Sterilize at 120°C 20 min in a closed vessel (i.e. a serum tube or bottle) with a liquid to air ratio of about 1:10. | |||||
2 After sterilization add from sterile stock solutions: | |||||
|
Trace element solution | 1.0 | ml | ||
|
MgCl2 x 6 H2O
(200.0 g/l) |
2.0 | ml | ||
3 (a white colloid will form which will rapidly dissolve after mixing) | |||||
|
Sodium thiosulfate pentahydrate
(4 M - 992 g/l) |
5.0 | ml | ||
|
Vitamin B12
(filter sterilized 100mg/l) |
1.0 | ml | ||
|
NaHCO3
(84.0 g/l) |
100.0 | ml | ||
4 Final pH is 8.0 - 8.5. | |||||
5 Incubate statically until growth occurs, then shake at 100 rpm. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
EDTA | 5 | mg | ||
|
FeSO4 x 7 H2O | 2 | mg | ||
|
ZnSO4 x 7 H2O | 100 | mg | ||
|
MnCl2 x 4 H2O | 30 | mg | ||
|
CoCl2 x 6 H2O | 200 | mg | ||
|
NiCl2 x 6 H2O | 20 | mg | ||
|
Na2MoO4 x 2 H2O | 30 | mg | ||
|
CuCl2 x 2 H2O | 10 | mg | ||
|
H3BO3 | 300 | mg | ||
|
Distilled water | 1000 | ml | ||
1 Final pH should be 3, add HCl if needed. Sterilization - 120°C 20 min. |