DEPC-treated water (RNase-free) recipe
RNase-free water for RNA work: treat with 0.1% DEPC, then autoclave to destroy the DEPC.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| DEPC (diethyl pyrocarbonate) | 1 mL | 0.1% (v/v) |
| Water (distilled) | to final volume |
Steps
- In a fume hood, add DEPC to the water and shake vigorously.
- Leave at room temperature for at least 2 h (usually overnight).
- Autoclave (121 °C, ≥15 min) to destroy residual DEPC (a faint fruity smell may remain).
Good to know
- Amine buffers such as Tris and HEPES react with DEPC — do not treat them directly; make them with DEPC water from RNase-free reagents.
⚠ Safety DEPC is a suspected carcinogen — handle in a fume hood with gloves.
FAQ
Is it the same as commercial nuclease-free water?
Same purpose; commercial water is filtered and tested without DEPC, which is safer for sensitive enzyme reactions.
This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.