RNA extraction (TRI Reagent and Direct-zol)

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

RNA extraction

1. Harvest cells by centrifugation at 1,000 × g for 5 min.
2. Resuspend the cell pellet in 800 μL TRI Reagent (Zymo Research).
3. Transfer to 2 mL Bead Ruptor Elite / Touch Micro-Organism Lysing Mix tubes (Omni).
4. Disrupt using a Precellys 24 Tissue Homogenizer (Bertin Instruments): 6,000 rpm, 15 s, 4 cycles, with a 2 min rest on ice between cycles.
5. Purify RNA using a Direct-zol RNA MiniPrep Kit (Zymo Research), per manufacturer’s instructions.
6. Measure RNA concentration and purity by NanoDrop.
7. Store RNA samples at −80 °C.