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| This
protocol is a simple procedure for growing up Paramecium in a monoxenic
culture of wheat grass media. It is cheap and easy to prepare.
Typically one will maintain a culture in a test tube and every three
to four days sterilely add about 1 mL of the inoculated Wheat grass
media to the culture. Transfers are accomplished by diluting
1 mL of cells with about 4 mLs of inoculated Wheat Grass in a new
sterile test tube.
Procedure for making Wheat Grass media. 1. Add 5 g of Wheat Grass to one liter of distilled water. Autoclave or boil for 20 minutes. 2. Filter the solution while still warm through two Whatman#42 filter papers. You may have to change filters. This is better done with some sort of vacuum. 3. Add 5mLs of Wheat Grass Buffer and 1 mL of Stigmasterol solution. Bring entire mixture back up to one liter volume if necessary. 4. Stir to mix and distribute into the appropriate size flasks. Autoclave. Procedure for preparing inoculated Wheat Grass media. Note: Uninoculated Wheat Grass media will kill a culture of Paramecia 1. Inoculate flask with single colony of Kelbsiella or Enterobacter. 2. Incubate at 37oC overnight. 3. Next day the flask is ready to use with your Paramecia culture. |
