457c .
MEDIUM WITH CHLOROACRYLIC ACID
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Equipment needed:
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Medium 457 | 1000 | ml | ||
|
|
3-chloroacrylic acid solution | 20 | ml | ||
| 1 Prepare medium 457. Make a stock solution of 4 g/l 3-chloroacrylic acid in water, neutralize and filter sterilize. Add 20 ml of this stock solution to 1 l of autoclaved medium. For better growth, 1 g/l glucose may be supplied additionally or instead of the chloroacrylic acid. | |||||
Main sol. 457 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 | 2.44 | g | ||
|
|
KH2PO4 | 1.52 | g | ||
|
|
(NH4)2SO4 | 0.50 | g | ||
|
|
MgSO4 x 7 H2O | 0.20 | g | ||
|
|
CaCl2 x 2 H2O | 0.05 | g | ||
|
|
Trace element solution SL-4 | 10.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 6.9. | |||||
| 2 Prepare a separate solution of the phosphates and autoclave separately. Combine the two solutions after cooling. | |||||
| 3 Rehydrate and cultivate lyophilized cells in complex medium (e.g. medium 1, 220 or 535). After this reactivation, cultivate in mineral medium 457 with the appropriate carbon source. | |||||
Main sol. 1 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Peptone | 5 | g | ||
|
|
Meat extract | 3 | g | ||
|
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Agar , for solid medium | 15 | g | ||
|
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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. | |||||
Trace element solution SL-4 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
EDTA | 0.5 | g |
|
|
FeSO4 x 7 H2O | 0.2 | g |
|
|
Trace element solution SL-6 | 100.0 | ml |
|
|
Distilled water | 900.0 | ml |
Trace element solution SL-6 #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
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 |