How To Avoid RNA Degradation

Avoiding RNA degradation is crucial for molecular biology experiments like RT-PCR, RNA sequencing, and Northern blotting. RNA is highly unstable and easily degraded by RNases, which are everywhere—even on your skin. Here’s how to prevent RNA degradation:

🧪 How To Avoid RNA Degradation

1. Use RNase-Free Materials

Only use certified RNase-free tubes, pipette tips, and water.
Autoclave materials or treat with DEPC (diethyl pyrocarbonate) to inactivate RNases.

2. Wear Gloves & Use Barrier Tips

RNases are abundant on your skin.
Always wear fresh gloves, and change them often.
Use filter (barrier) pipette tips to avoid cross-contamination.

3. Clean Work Area with RNase Decontamination Solutions

Wipe down benches, pipettes, and equipment with solutions like RNaseZap or 70% ethanol + DEPC-treated water.

4. Work Quickly and on Ice

Perform all steps quickly and keep samples on ice to slow down RNase activity.
Pre-chill reagents and centrifuges.

5. Use RNase Inhibitors

Add RNase inhibitors (like RNasin or Protector RNase Inhibitor) to your reactions if applicable.

6. Properly Store RNA Samples

Store RNA in:
- RNase-free water or TE buffer (pH ~7.5)
- Aliquots to avoid freeze-thaw cycles
- -80°C for long-term storage
- Add formamide for extra stability in some protocols

7. Avoid Using Contaminated Reagents

Always use freshly opened, RNase-free reagents.
Never touch reagent tips or tube interiors.

8. Use Commercial RNA Stabilization Reagents

For tissue or blood samples, use reagents like RNAlater to stabilize RNA immediately upon collection.

Summary Table

Step	Why It Helps
RNase-free consumables	Prevents contamination
Gloves and barrier tips	Avoids skin-derived RNase transfer
RNaseZap or ethanol cleaning	Kills RNases on surfaces
Work quickly/on ice	Reduces RNase activity
Use RNase inhibitors	Protects RNA during handling
Store properly (-80°C, aliquots)	Prevents degradation over time
Use RNAlater/stabilizers	Preserves RNA immediately