title: RNA extraction from mammalian cells — protocol v1 project: research-lab-template tags:

• protocol
• rna
• wet-lab
• safety date: 2026-04-07

RNA extraction from mammalian cells — protocol v1

Status: Active | Owner: Lab manager | Revision: v1 (template)

Scope

Total RNA from 1e6–5e6 adherent or suspension mammalian cells using column-based cleanup. For BSL-2 material, complete institutional training before use.

Materials

RNA kit (log vendor lot), 70/100% ethanol, optional on-column DNase per kit insert, β-mercaptoethanol (hood only; toxic).

Steps (summary)

Harvest → PBS wash → pellet; lyse with fresh reducer (vortex 10s, 5 min RT); 70% ethanol bind; column washes per insert; optional DNase for qPCR-grade RNA; elute 30 µL water; A260/280 and RIN if sequencing.

Safety

• β-mercaptoethanol: use in chemical hood; avoid skin contact.
• Phenol-chloroform variants (if substituted): only in designated hood with training.

QC & revisions

Target A260/280 ~1.9–2.1; store at −80°C if not used the same day. v1: initial Knowtation template structure.