Wes wrote: ↑Wed Sep 16, 2020 6:02 pm
PROTOCOL
Spirostomum, Stentor
1. Medium ingredients should be added in the sequence listed. If the name of a company is listed
with a specific component, the medium component should be obtained from the company to
assure growth.
2. Medium formulation:
Soil/H20 Biphasic Medium [1]
Rich Organic Soil (Air-dried) .......... 125.0 g
Glass distilled H20 ...................... 875.0 ml
3. Place soil in the bottom of a flask and gently overlay with the glass distilled H20, etc.
4. Autoclave at 121 0 C for 15 min. Allow to cool overnight.
5. Repeat step 4.
6. In advance place wheat kernels in a 20 x 125 mm screw-capped test tube to a depth of
approximately 5 cm. Add a few drops of glass distilled H20 and with the cap loosened one half
turn, autoclave for 30 min at 121˚ C. Place the tube in a 70˚ C incubator overnight to remove
any moisture. Allow the tube to cool to room temperature, screw the cap on tight and store
refrigerated (5-9˚ C) until needed. Wheat kernels may be obtained from a grocery store.
7. Allow the flask prepared in step 5 to cool, swirl vigorously and transfer 200 ml to an 11 cm
diameter biological finger bowl. Add one autoclaved wheat kernel.
8. Inoculate 1 ml of a growing culture into medium prepared in step 7
9. Place another empty finger bowl on top of the culture to reduce moisture loss and incubate at
25° C.
10.Repeat steps 8-10 at monthly intervals.
9. Place another empty finger bowl on top of the culture to reduce moisture loss and incubate at
25° C.
10.Repeat steps 8-10 at monthly intervals.
