Brief introduction to the importance of RNA
RNA (ribonucleic acids) are important macromolecules that come in many varieties and play more roles across all living organisms. Some predominant RNA are rRNA, tRNA and mRNA. tRNA is part of the process of translating proteins, rRNA is ribosomal RNA, and mRNA is messenger RNA which carries transcription information. RNA biology is an ever-growing field of research. RNA has even become a popular tool, such as the mRNA COVID-19 vaccine or CRISPR technologies. Researchers are even finding more non-coding RNA with important biological functions.
Considerations for working with RNA purification kits
There are many types of available RNA purification kits. Different kits will address potential problems or downstream applications for various RNA purification protocols. The biggest challenge to be aware of when working with RNA is the potential to degrade your material from RNases present all over your skin. When working with RNA, it is important to clean the working space with a cleaner that will kill RNases, wear clean gloves, and avoid sneezing or talking a lot near the sample. If your starting material is tissue, you will want to consider the best lysis protocol to release your cell contents. You can read more about this step here.
RNA purification kits
Spin Column RNA purification kit
There are many types of RNA purification kits that are slightly optimized towards various experiments. Some kits are specific for purifying RNA from fungi, bacteria, plants, blood etc. Some kits contain buffers and columns to perform the lysis steps and go straight into purification steps. The basic principle of the first type of RNA purification kit technology is the spin column. The kit will contain all the buffers you need, lysis buffer, binding buffer, and wash buffers. The RNA purification kit will also come with spin columns which have some proprietary version of a silica based matrix that will bind RNA in the correct pH and salt environment. There will be a protocol for adding a sample, and centrifuging at each step of the process from binding to washing to eluting. These kits often come in eppendorf tube sizes to 96 well plates, called “mini”, “midi” or “Max” prep kits. The main disadvantage of spin column techniques is their generally low yield and more complicated procedures.
Magnetic Bead RNA purification kit
Magnetic bead RNA purification is a simple way to isolate RNA from your sample. This kit also requires buffers for each step, binding, washing, eluting. The technology that coats the surface of the bead is similar to the spin-columns, combining a silica bead surface with the correct buffer environment for RNA binding. Magnetic bead separation has the benefit of quick and safe separation by placing the sample container in a magnetic separator, instead of using a centrifuge. Modern magnetic separators come in many sizes and allow you to monitor the separation on the computer. This can speed the optimization protocol when creating a method in your lab for RNA isolation. Due to the reliability of the magnetic separator, the yield from magnetic bead RNA purification is higher than for spin-columns.
Benefits of RNA purification kits
Purifying RNA is a widely used technique to prepare samples for RNA-seq or for diagnostic purposes. The optimization of many types of RNA purification kits has allowed research labs and clinics to increase their utilization of the amazing nucleic acid. If you are interested in bringing an RNA purification protocol to your lab, you can get started on designing a protocol here.
