Carolina C-FERN Spores - Manual (Page 4)

4
Temperature
• The optimum temperature for spore germination and gametophytic
development is 28°C (82°F), but results are good at any temperature
within the range of 26–30°C (79–86°F).
• Lower temperatures will substantially alter development.
• Monitor and record temperatures inside the Growth Pod/Culture Dome
daily.
• Adjust the distance between the light and the Growth Pod/Culture Dome
to control temperature. Once a suitable temperature is achieved, the
height of the light should remain constant.
Troubleshooting—Condensation
Condensation on petri dish lids may occur if there is excessive temperature
variation. Moderate amounts of condensation are not a problem, but
excessive condensation may result in an uncontrolled release of
spermatozoids. If condensation becomes a problem, grow cultures upside
down once the sowing water has been absorbed in the medium.
Preparing the Medium
Bottled Basic C-Fern ® Medium (RN-15-6780, 160 mL; RN-15-6781, 400 mL)
1. Melt the medium.
(To speed up this process, shake the bottle to break up the medium.)
• Loosen the cap on the container.
• Place the container in a hot water bath such that the water level is
just above the level of the medium.
• Do not submerge the container!
2. Pour the medium into the petri dishes.
• Medium should be poured in a clean area free from drafts and
traffic.
• Wipe down the area with 70% ethanol or isopropanol.
• Open a sleeve of 60-mm petri dishes.
• Check that the medium is completely melted.
• Gently swirl the medium to ensure it is mixed.
• Tilt the lid of a petri dish just enough to permit pouring.
• Fill the dish about 3 /
4
full (about 15 mL per dish). Note: Do not
underfill the dishes.