protocols-io/TRIzol RNA Extraction

This protocol describes the isolation of total RNA from mammalian cells and tissue samples using TRIzol reagent. TRIzol is a monophasic solution of phenol and guanidine isothiocyanate that simultaneously lyses cells and inhibits RNases, enabling high-quality RNA isolation from a variety of biological samples.

Overview

The procedure involves cell lysis in TRIzol, phase separation with chloroform, RNA precipitation with isopropanol, and washing with ethanol. When performed correctly, yields RNA with RIN values above 8.0, suitable for downstream applications including RT-qPCR, RNA-seq, and microarray analysis.

Key considerations

• Sample input: Works with adherent cells (directly in culture dish), cell pellets, and tissue (up to 100 mg per mL TRIzol)
• RNase prevention: Use RNase-free consumables throughout; work on ice where indicated
• Yield expectations: ~5–10 µg total RNA per 1 million cells; tissue yields vary by type
• Quality control: Assess RNA integrity via Bioanalyzer or TapeStation before downstream use