MuLTEERatcliff Lab · Georgia Tech

Protocols / Freezer stocks

Reviving strains from frozen stock

Freezer stocksBy Various authorsVersion 1 · updated 2026-10-07

Bringing strains back from −80 °C stocks and re-acclimating them to full cluster size with settling selection.

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Adapted from the lab compendium Snowflake Yeast Protocols — Ratcliff Lab / MuLTEE (V2).

  1. Using a P10 pipette tip, scrape a small amount of ice directly from the surface of the frozen stock.
  2. Inoculate the scraped material into 10 mL fresh YPD.
  3. Grow out following the standard growth protocol (Culturing & settling selection › Daily growth and settling selection › Growth) before beginning any re-acclimation or further propagation.
  4. Go through 3–5 rounds of settling selection (Culturing & settling selection › Daily growth and settling selection › Settling selection) to encourage growth back to full size. Taking 15 μL transfers instead of 50 μL transfers from settled culture sometimes accelerates this process.