Handling RNA samples requires careful techniques to prevent degradation, primarily from RNases, enzymes that break down RNA. Following these guidelines will maintain the integrity of your RNA for experiments. Here's how to handle RNA samples:
Essential Precautions for RNA Handling
Maintaining a Clean Work Environment
- Use Sterile Technique: This is fundamental to preventing RNase contamination.
- Wear clean gloves and a fresh lab coat to avoid introducing RNases from skin.
- Keep tubes and bottles sealed tightly to protect samples.
- Work in a clean, dedicated RNase-free area.
- Use RNase-Free Supplies: This is critical to prevent sample degradation.
- Utilize RNase-free tubes, tips, and other consumables.
- Use RNase-free solutions and reagents in your experiments.
Table of Important RNA Handling Practices:
| Practice | Why? |
|---|---|
| Wear clean gloves & lab coat | Prevents RNase contamination from skin. |
| Keep tubes and bottles closed | Protects sample from environmental RNases. |
| Use an RNase-free workspace | Minimizes risk of RNase exposure. |
| Use RNase-free consumables | Avoids introducing RNase through lab supplies. |
| Use RNase-free reagents | Prevents introducing RNases through chemicals. |
Detailed Explanation of Key Practices
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Personal Protection:
- Always wear disposable gloves when handling RNA. Change them frequently, especially if you touch anything that might be contaminated.
- Put on a clean lab coat. Dedicate one lab coat for RNA work.
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Work Area Preparation:
- Clean your workspace with RNase decontamination solution before starting.
- Use a clean bench area that is specifically designated for RNA work.
- If possible, work in a laminar flow hood to maintain a sterile environment.
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Consumables and Reagents:
- Make sure all tubes, pipette tips, and other consumables are certified RNase-free.
- Purchase reagents specifically labeled as RNase-free. If you prepare your solutions, use DEPC (diethylpyrocarbonate)-treated water.
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Handling Samples:
- Keep RNA samples on ice or in a -80°C freezer whenever they are not actively being used.
- Minimize the number of freeze-thaw cycles. Aliquot the RNA into smaller volumes to avoid repeated thawing.
- Avoid vortexing RNA solutions. Instead, gently mix by inverting the tube.
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Best Practices for Long-Term Storage:
- Store RNA at -80°C for extended periods.
- Precipitate RNA with ethanol and store as a pellet for even more long-term storage.
By following these steps and paying close attention to detail, you can ensure the integrity of your RNA samples, which is critical for successful molecular biology research.
