Carolina C-FERN Spores - Manual (Page 6)

6
Preparing Spores
Tap the vial on a hard surface so that the spores are at the bottom of the
vial before opening it.
Presterilized Spores
• Loosen the cap.
• Transfer the appropriate amount of sterile distilled water to the spore
vial with a sterile pipet (the standard presterilized spore unit will sow
about 35 dishes at a density of 300+ spores per plate when 4 mL of
sterile distilled water is added).
• Do not return any water to the sterile water bottle.
• Wet the spores completely by firmly tightening the cap and inverting the
vial 2–3 times.
• With the cap on, check the bottom of the vial to ensure that all spores
have been suspended.
• Return the vial to the holder and loosen the cap.
Unsterilized Spores
Sterilize spores according to the following procedure:
• Weigh the spores on glassine weigh paper and transfer them to a sterile,
conical, 5-mL tube. (10 mg will inoculate 35 dishes.)
• Presoak the spores by covering them with 1–2 mL of sterile distilled
water for 15 minutes to 24 hours.
• Remove the presoak water:
a. Insert a sterile pipet into a conical tube.
b. Suspend the spores by bubbling a small
amount of air into the water.
c. Gently, but securely, seat the pipet tip at the
base of the tube.
d. Squeeze the bulb to force air out of the
pipet.
e. Release the bulb to draw water into the
pipet while spores collect around the tip.
• Surface sterilize the spores:
a. Suspend the spores in 1–2 mL of 0.875% sodium hypochlorite.
b. Rinse down the lip and sides of the spore vial with sodium
hypochlorite solution.