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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.