Although proper handling is essential when working with DNA and RNA, the latter requires extra care because it is single-stranded polynucleotide, which makes it vulnerable to debasement by base- or enzyme-catalyzed hydrolysis. Another reason why extra caution is needed while working with RNA is, it is chemically quite unstable and presence of RNases is extremely high. We know that DNases call for metal ions for activity, which is not the case with RNases as they can maintain activity even in the absence of metal ions. Autoclaving or lengthy boiling seem to have no effect on their activity.
The problem with RNases is, they are literally everywhere. You can find them on the hands of laboratory workers and in airborne microorganisms. That is why special care needs to be taken while working with RNA. Before using all reagents and equipment, make sure they are specially treated to inactive RNases. In this blog, we will share with you some key tips that will help you keep your laboratory environment free of RNases. What are those tips? Let’s find out.
It is essential to wear gloves when you are working with RNA. And once you’ve got your gloves on, make sure you do not touch any contaminated surface and equipment. All the reagents might be fully decontaminated, but keep in mind that RNases can contaminate it anytime through unfiltered air or ungloved hands.
We will strongly recommend you to use sterile, disposable plasticware as they do not call for any special treatment and are known for being RNase-free. As far as cleaning the electrophoresis tanks for RNA analysis is concerned, you can use a solution of SDS (1%) to wipe them and rinse with water. It is advisable to rinse them with absolute ethanol and then soak them for ten minutes in 3% H2O2.
It would be good to get reagents that are RNase-free. Do not forget to divide reagents used for RNA work from regents used for general use in the laboratory. Barring Tris buffers, make sure all other solutions are treated with 0.1% DEPC all-night at room temperature and then autoclaved. Autoclaving is required because it is known for hydrolyzing and destroying unreacted DEPC and DMPC.
Apart from doing the above-mentioned things, you should also fix an area purely for RNA work. Use commercially available RNase inactivating agents to clean surfaces of benches and glassware. Get automated RNA clean-up kit from AMD Biotech. This kit can be used for PCR and RT-PCR reactions, in vitro RNA synthesis, transfection for RNAi experiments, and much more. The kit contains paramagnetic particles and reagents for high-throughput purification of RNA or cDNA.