YEPAD Freezing

A Dohlman Lab Protocol

Yeast strains, transformed or untransformed, can be maintained as colonies on solid media at 4° and restreaking every 2 to 4 weeks. Alternatively, strains may be stored at -80 indefinitely. This is preferable in that it reduces the likelihood of accumulating spontaneous mutations.

To make frozen stocks of yeast strains: -Use sterile technique and sterile solutions throughout this method.-

  1. Grow a starter culture at 30 with shaking (250 rpm) until it reaches saturation.
  2. In a 1.8 mL cryotube, mix 0.5 mL of the saturated culture with 0.5 mL of YPD containing 20% glycerol.
  3. Flash freeze the tube in liquid nitrogen and store at -80.
  4. To use, chip out a few pieces of the frozen stock using a sterile pipette tip or sterile toothpick and streak onto a plate containing the appropriate solid medium.
  5. Allow the yeast to grow at 30 until colonies appear (2-6 days).