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(Source: W. Driever and C. Boggs from Zebrafish Book 5th Edition)

Preparation of paramecia culture medium

The classical hay infusions are often unreliable (and tend to stink!) and some people have mixed results with mung beans or wheat.  The following is a modification of an axenic method for growing paramecia (Schöne­feld et al. J. Protozool. 33:222).

Schönefeld's Medium for Growing Paramecia:

Component

Concentration

Source

Powdered Skim Milk

8.5 g/l

Grocery store

Ribonucleic Acid

1.0 g/l

Sigma R-6625

MgSO4

0.5 g/l

Sigma M-2643

Phosphatidylcholine

250 mg/l

Sigma P-5638

Stock Solution I

5.0 ml/l

 

Stock Solution II

2.5 ml/l

 



1.   Prepare the solution of primary components.  Make sure the dry ingredients are well mixed before adding them with stirring to the water or the ribonucleic acid and the phosphatidylcholine will lump together and not dissolve.  This solution can be heated slightly to help dissolve small lumps.  Then add Stock Solutions I and II.

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.

Paramecium Medium Stock Solution II:

 

g/l in EtOH

Source

Stigmasterol

2.0

Sigma S-6126

Lipoic Acid(Dl‑6,8‑thioctic acid)

0.02

Sigma T-1395

Palmitic Acid

3.0

Sigma P-5917

Stearic Acid

2.0

Sigma S-4751

Oleic Acid

0.4

Sigma 0-4379

Linoleic Acid

0.2

Sigma L-1376

Linolenic Acid

0.06

Sigma L-2376


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.