462 .
MINERAL MEDIUM (BRUNNER) WITH VITAMINS
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* Medium for Hyphomicrobium sp. DSM 18203
Plus 2g/l methanol and dichlormethane via atmosphere
Growth conditions: 28 °C
Equipment needed:
Autoclave
Final pH:
6.9
Select a strain (optional):
ml
| 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 | ||
|
|
Vitamin solution
(Schlegel) |
2.50 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 6.9. | |||||
| 2 Rehydrate and cultivate lyophilized cells in the complex medium recommended for the specific strain (e.g. medium 1, 220 or 535). After this reactivation, cultivate in mineral medium 462 with the appropriate carbon source. | |||||
|
|
Medium 1 | 1000.00 | ml | ||
|
|
Medium 220 | 1000.00 | ml | ||
|
|
Medium 535 | 1000.00 | ml | ||
|
|
Methanol | 2.00 | g/l | ||
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 |
Vitamin solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
p-Aminobenzoate | 1.0 | mg |
|
|
Biotin | 0.2 | mg |
|
|
Nicotinic acid | 2.0 | mg |
|
|
Thiamine-HCl x 2 H2O | 1.0 | mg |
|
|
Calcium pantothenate | 0.5 | mg |
|
|
Pyridoxamine | 5.0 | mg |
|
|
Vitamin B12 | 2.0 | mg |
|
|
Distilled water | 100.0 | ml |
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. | |||||