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(Source: W. Driever and C. Boggs from Zebrafish Book 5th Edition)
Preparation of paramecia culture medium
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Paramecium Medium Stock Solution I:
| g/l in dH2O | Source |
|---|---|---|
Calcium pantothenate | 1.0 | Sigma P-2250 |
Nicotinamide | 1.0 | Sigma N-3376 |
Riboflavin | 1.0 | Sigma R-9881 |
Pyridoxamine HCL | 1.16 | Sigma P-9158 |
Folic Acid | 0.5 | Sigma F-7876 |
Thiamine HCL | 3.0 | Sigma T-4625 |
Biotin | 0.00125 | Sigma B-463 |
Make 100x stock. Store at -20° C; the riboflavin is dispersed through-out the solution and will settle to the bottom. Shake well before using.
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Blow nitrogen gas over the solution and store at -20°C.
2. Adjust the pH of the medium to 7.2 at room temperature.
3. Autoclave for 15 minutes in 500 ml aliquots (Some coagulation may occur, which is ok, but if too much of the milk precipitates, the paramecia will not grow).
4. Test whether the medium is sterile and store in a refrigerator for up to several months.