Freezing
- glycerol (most fungi an bacteria do well in 10%, some need 20%) or mineral oil (messy)
- Put three agar plugs (or just chunks of mycelium, or one big chunk, whatever is easier) from each plate into its respective storage vial. Agar plugs (cubes with fungus) or pieces of mycelium can be cut with scalpel; make sure that scalpel is perfectly sterilized after each culture. Take the growing edge of the fungus, or a whole growing colony, not the old crusty center.
- If you are using minislants – just pour the sterile 20% glycerol in the minislant tube
- Place in Mr. Frosty (blue/white container on top of the fridge). Fill up the bottom compartment with isopropyl alcohol (in the chemicals storage). Put in -80C freezer. It will freeze slowly, 1C per minute.
- Record in the Scolytos Database!!! Each isolate will get genus name “fungus” and its unique Species name will be its Isolate name from the isolations database. In “count”, record 1.
Reviving
Prepare:
- vials with 1mL PBS. (Label them with numbers corresponding to frozen samples)
- equal number of PDA plates (label these with regular database numbers)
- sterilizer, scalpel
1. Take out sample. 2. Cut gel disk inside the tube with carefuly sterilized scalpel. 3. Use only one half (other half stays). 4. Put half-disk in vial with PBS, shake briefly to rinse off surplus glycerol or mineral oil, and put on PDA plate.
