465f .
MINERAL MEDIUM WITH 3-AMINOPHENOL
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Equipment needed:
Autoclave
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Final pH:
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Medium 465 | 1000 | ml | ||
|
|
3-Aminophenol
(10 g/l) |
10 | ml | ||
| 1 Prepare mineral medium 465. Prepare a stock solution of 10 g/l 3-aminophenol by adding boiling water to the crystals, stir the solution, cool and sterilize by filtration. Add 10 ml of this stock solution to 1 l of autoclaved and cooled medium 465. | |||||
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 | ||
|
|
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. | |||||
Main sol. 1 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Peptone | 5 | g | ||
|
|
Meat extract | 3 | g | ||
|
|
Agar , for solid medium | 15 | g | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH to 7.0. | |||||
| 2 For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation. | |||||
Main sol. 220 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Casein peptone | 15 | g | ||
|
|
Soy peptone | 5 | g | ||
|
|
NaCl | 5 | g | ||
|
|
Agar | 15 | g | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH to 7.3. Medium is identical with Tryptone Soya Agar (Oxoid CM 131). | |||||
Main sol. 535 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Trypticase soy broth
(BBL 11768, Oxoid CM129 or Merck 5459) |
30 | g | ||
|
|
Agar | 20 | g | ||
|
|
Distilled water | 1000 | ml | ||
| 1 pH 7.3 | |||||
| 2 Autoclave at 121°C for 15 min. | |||||
Main sol. 830 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract | 0.50 | g | ||
|
|
Proteose peptone
(Difco no. 3) |
0.50 | g | ||
|
|
Casamino acids | 0.50 | g | ||
|
|
Glucose | 0.50 | g | ||
|
|
Starch
(soluble) |
0.50 | g | ||
|
|
Na-pyruvate | 0.30 | g | ||
|
|
K2HPO4 | 0.30 | g | ||
|
|
MgSO4 x 7 H2O | 0.05 | g | ||
|
|
Agar | 15.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Final pH 7.2; adjust with crystalline K2HPO4 or KH2PO4 before adding agar. Add agar, heat medium to boiling to dissolve agar, and autoclave for 15 min at 121°C. | |||||