Preventing RNAse Contamination

How is RNAse contamination in RNA based experiments prevented?

RNAse contamination is a common problem in RNA-based experiments that can lead to degradation and loss of RNA samples. To prevent RNAse contamination, several precautions and strategies can be implemented:

1. Use dedicated equipment: Ensure that all equipment, such as pipettes, microcentrifuges, and water baths, are designated solely for RNA work. This prevents cross-contamination from RNAse-rich environments and samples

2. Wear gloves and lab coats: Always wear gloves and lab coats while handling RNA samples to avoid introducing RNAse enzymes from your hands or other external sources

3. Practice good laboratory techniques: Follow proper aseptic techniques while working with RNA samples, including using sterile RNA-free tubes, filter tips for pipetting, and clean bench surfaces that have been treated with RNase inhibitors (e. g. , RNase AWAY)

4. RNase-free reagents: Use RNA-grade reagents that are certified as RNase-free. This includes using RNase-free water, buffers, and chemicals throughout the entire procedure. It is essential to verify the quality of commercial reagents and confirm that they are specifically designed for RNA work

5. Clean workspace: Regularly decontaminate your working surface and equipment with RNase decontaminating agents (e. g. , 0. 1% DEPC-treated water) to remove any residual RNase activity

6. Pre-treat solutions and equipment: Before performing RNA work, treat all solutions, tubes, and plasticware by autoclaving or baking at high temperatures (300°C) to inactivate any contaminating RNases

7. UV irradiation: UV irradiation can be used to inactivate RNases on the surface of equipment or when working in a UV-safe cabinet. However, caution must be taken as UV exposure may lead to RNA degradation if not controlled properly

8. RNase inhibitors: Incorporate RNase inhibitors, such as RNasin or SUPERase-In, into the reaction mixtures to provide an additional layer of protection against any residual RNAse contamination

9. Store RNAse-free: Store all RNA samples and reagents in RNase-free conditions, such as in low-bind tubes, RNAse-free containers, or under appropriate low-temperature conditions, to prevent degradation

10. Work quickly: Complete the RNA handling steps as quickly as possible to minimize the exposure time of RNA samples to the environment, reducing the chances of RNAse contamination

It is important to note that even with the implementation of these preventive measures, complete elimination of RNAse contamination is challenging. Therefore, it is advisable to frequently monitor the quality and integrity of RNA samples using techniques like gel electrophoresis or real-time PCR to ensure reliable results in RNA-based experiments

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