465i .
MINERAL MEDIUM WITH ATRAZIN
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Equipment needed:
Autoclave
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
2-chloro-4(ethylamino)-6-(isopropylamino)-1,3,5-triazine
, adjust if required
(suspend 10 mg/ml in methanol) |
100 | mg/l | ||
|
|
Medium 465 | 1000 | ml | ||
| 1 Suspend atrazine (2-chloro-4(ethylamino)-6-(isopropylamino)-1,3,5-triazine) in methanol (10 mg/ml). Shortly sonicate to reduce particle size. Add stock suspension to medium 465 without nitrogen source (omit ammonium sulphate and calcium nitrate), replace calcium nitrate by calcium chloride) but with 1 g/l glucose as the carbon source. Final concentration of atrazine is 50-100 mg/l. | |||||
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. | |||||