Small culture slants have the advantage of easy storage, integration with our database system, and safe shipping to collaborators.
- Sterilize 2mL cryotubes by autoclaving or prolonged exposure to UV light.
- Under a biosafety hood, arrange the open tubes in a rack so that the open end of the tubes face almost horizontally (about 15° ).
- Expose the tubes to UV light again if necessary to ensure they are not contaminated.
- Add about 1mL of autoclaved media (usually PDA) to the bottom of the tube.
- Let the media harden and cool UNDER AN ANGLE in the hood before adding the tube caps.
- Store in the fridge.
Following this, you may culture the fungus on the slant media, label it with a scolytos vial number, and place it in the incubator, slightly open with parafilm, to grow. After the media has a culture, you may add 10-20% glycerol to fill the vial, close it completely and use the “Mr. Freeze” jar to slowly bring it down to freezing in the -20C. It can then be moved to the -80C for long term storage.
For sending the tubes to collaborators:
- Keep the culture for a couple days before shipping to make sure the fungus is growing and isn’t contaminated.
- Wrap the lid-tube joint with parafilm, so that the lid doesn’t get loose.
- Tape the tube to a piece of paper so that it doesn’t roll around in the envelope during shipping.
- Make sure it is sufficiently padded and that the tube and envelope are labeled.
- Ship IMMEDIATELY: there is very little oxygen for the fungus to breathe.