Protocol:CULTURE STORAGE (10% GLYCEROL)
Freezing- Cryoprotection
Need: 2 ml cryotubes (sterile)- 2 per isolate, holding rack, sterile pipet tips ( 1 ml volume ), P 1000 pipetman, sterile 10% glycerol, freezer storage box, styrofoam container with ice, 1 cm cork borer.
- Place 1 ml 10% Glycerol in each tube .
- Cut plugs with cork borer from a 1-2 week old culture. Use a fresh plate ( never opened) and take plugs 1 cm from outer edge of mycelia.
- Stack 5 plugs in each tube ( make sure Glycerol covers everything- remove 1 plug if necessary).
- Label tubes with isolate number and date and put on ice.
- Transfer tubes to freezer box and place immediately in the - 80 C freezer for 30 min. ( up to 24 hrs. if necessary).
- Put tubes in assigned locations in - 135 C freezer.
