Protecting RNA involves carefully preventing its degradation, primarily by avoiding contamination with RNases, which are enzymes that break down RNA.
Understanding the Threat: RNases
RNA is notoriously fragile and susceptible to degradation by enzymes called ribonucleases (RNases). These enzymes are abundant and resilient, found on surfaces, in the environment, and even on our skin. The key to protecting your RNA samples is to eliminate or inhibit these RNases.
Practical Steps for RNA Protection
Based on best practices in molecular biology laboratories and the provided reference, here are essential methods to safeguard your RNA:
1. Prevent Contamination from Skin
Your skin is a significant source of RNases. To avoid introducing these enzymes into your samples or workspace:
- Always wear gloves when handling RNA, reagents, tubes, and lab equipment used for RNA work.
- Change gloves frequently, especially after touching non-sterile surfaces or before handling critical reagents or samples.
2. Use RNase-Free Reagents and Water
Ensure that all materials coming into contact with your RNA are free from RNase contamination:
- Use certified reagents specifically labeled as RNase-free.
- Crucially, use high quality water, such as nuclease-free or DEPC-treated water, for preparing solutions and diluting samples. Standard lab water is often contaminated with RNases.
3. Employ RNase Inhibitors
Even with stringent sterile techniques, a small amount of RNase contamination can be detrimental. RNase inhibitors are proteins or chemicals that bind to and inactivate RNases.
- Use an RNase inhibitor, such as RiboLockTM RNase Inhibitor (as mentioned in the reference) or similar products, in your reactions or storage buffers to actively protect the template RNA or synthesized RNA.
Summary of RNA Protection Methods
Here's a quick overview of the key strategies:
| Protection Method | Why It Works | Practical Action |
|---|---|---|
| Prevent RNase Contact | Skin is a major source of RNases | Wear and frequently change gloves. |
| Use Clean Reagents | Reagents and water can harbor RNases | Use certified RNase-free reagents and nuclease-free water. |
| Inhibit RNase Activity | Inactivates residual RNases in the environment | Add an RNase inhibitor to reactions and buffers. |
By implementing these preventative measures, you can significantly reduce the risk of RNA degradation and ensure the integrity of your samples for downstream applications.
