465c .
MINERAL MEDIUM WITH DICHLOROMETHANE
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Equipment needed:
Autoclave
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Medium 465 | 1000 | ml | ||
| 1 Prepare medium 465 (addition of 50 mg/l bromothymol blue as pH-indicator is recommended) and autoclave. Grow the cells in medium 465 with methanol supplied via incubation atmosphere (up to 10 ml methanol per liter medium). Incubate these adapted cells in medium 465 and consecutively feed small amounts of dichloromethane (toxic! 2 mmol per liter or less) via incubation atmosphere. Readjust pH of the culture with sterile 1 M NaOH if necessary. | |||||
Main sol. 465 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 x 2 H2O | 3.50 | g | ||
|
|
KH2PO4 | 1.00 | g | ||
|
|
(NH4)2SO4 | 0.50 | g | ||
|
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
|
Ca(NO3)2 x 4 H2O | 0.05 | g | ||
|
|
Trace element solution SL-4 | 1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 pH 7.25 | |||||
| 2 Rehydrate and cultivate lyophilized cells in the complex medium recommended for the specific strain (e.g. medium 1, 220, 535 or 830). After this reactivation, cultivate on mineral medium with the appropriate carbon source. | |||||
Trace element solution SL-4 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 0.50 | g | ||
|
|
FeSO4 x 7 H2O | 0.20 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
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MnCl2 x 4 H2O | 0.03 | g | ||
|
|
H3BO3 | 0.30 | g | ||
|
|
CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.03 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. | |||||