465d .
MINERAL MEDIUM WITH BENZYLCYANIDE AS NITROGEN SOURCE
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* Medium for Rhodococcus erythropolis DSM 9675
Instead of 1.8 g/l glucose add 1.6 g/l sodium succinate from a sterile solution.
Growth conditions: 28 °C
Equipment needed:
Autoclave
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Main sol. 465 (modified) | 1000.0 | ml | ||
| Glucose* | 1.8 | g/l | |||
|
|
Benzylcyanide | 120.0 | mg/l | ||
|
|
Sodium succinate | 1.6 | g/l | ||
| 1 Prepare and autoclave medium 465 without ammonium sulfate. Add 120 mg/l benzylcyanide (not sterilized) to the incubation atmosphere or directly to the medium. | |||||
Main sol. 465 (modified) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 x 2 H2O | 3.50 | g | ||
|
|
KH2PO4 | 1.00 | g | ||
|
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MgCl2 x 6 H2O | 0.10 | g | ||
|
|
Ca(NO3)2 x 4 H2O | 0.05 | g | ||
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Trace element solution SL-4 | 1.00 | ml | ||
|
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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 | ||
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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. | |||||