The purification of RNA is important for research laboratories and clinics. Throughout the COVID19 pandemic, the purification of RNA has been in the news because the rate of RNA purification defines how quickly testing for COVID19 can be done. RNA is now in the news again, mRNA specifically, as major companies have developed mRNA vaccines for COVID19. In brief, the vaccine is an injection that sends lipid coated mRNA into cells. In cells, the mRNA is translated into the spike protein of SARS-CoV-2. Companies have shown robust responses to the vaccines as the creation of the protein in cells elicits a good amount of response from the correct immune cells.
mRNA purification kit, how they work
There are various types of kits commercially available for mRNA purification. The main principle of these kits for mRNA capture is the use of mRNA polyadenylation, a process at the end of transcription which creates a string of adenosine nucleic acid molecules at the end of each mRNA transcript. Most kits are now magnetic bead kits.
- In one type of kit, streptavidin will be conjugated to magnetic beads, while biotin is linked to oligo-dT molecules in solution. First, mRNA are mixed with the oligo-dT-biotin molecules, the polyA tail binds to the oligo-dT chain. Second, the mRNA bound biotin molecules are mixed with the pre-conjugated magnetic beads where biotin binds readily to streptavidin. Lastly, in the presence of a modern magnetic separator, the mRNA molecules will safely and quickly separate from their mixture by the magnetic force acting on the beads. After a few seconds, the magnetic beads with the mRNA are moved to the walls of the container and retained by the magnetic force, when the rest of the mixture can be discarded simply pumping-out the supernatant.
- In another type of kit, oligo-dT is pre-conjugated directly to magnetic beads. This process has fewer steps. One simply has to add the total RNA mixture to solution with oligo-dT pre-conjugated magnetic beads, and only the polyA tails of mRNA will bind the magnetic beads. As mentioned before, a modern magnetic separator is used to keep the mRNA in the container, while the rest of the mixture can be discarded.
mRNA purification kit, choosing a magnetic separator
Purifying mRNA is vital to research, clinical testing, and creating modern vaccines. These environments necessitate quick purification, and the ability to scale up production of mRNA purification. Modern biomagnetic separation systems offer the capacity to scale up mRNA purification, while keeping the process simple and fast. These new devices are made to create a constant magnetic force to ensure that samples are separated safely and steadily, as well as quickly. The constant value of the magnetic force allows characterization and validation of the magnetic separation process, which can then be replicated at different volumes. With these biomagnetic separation systems it is straightforward to scale up the process up to tens of litres.
mRNA purification kit, considerations for your purification
When working with RNA, an RNase free environment must be maintained. RNases are all over our bodies, and have the potential to contaminate RNA samples and degrade them. There are solutions that degrade RNases that are liberally sprayed on any surface or tool used for mRNA purification. Gloves are changed often, and ideally there is no talking near samples. The danger of RNA degradation is a consideration that makes the use of magnetic beads for mRNA purification so popular. The procedure is simple and consistent, one simply has to put their container in a magnetic separator, and remove solution.