Speaking of viruses, take the new coronavirus COVID-19: SARS-coV-2 that we now hate as an example. Its genetic material is RNA. If the SARS-CoV-2 pandemic gives us any inspiration, the study of RNA modification is now more important than ever. Does this mean studying the viral RNA itself to learn more about how different changes and epigenetic changes make these viruses more resilient and infectious, or studying RNA collected from cells and tissues to understand these differences The effect of pathogens on host cell function is almost endless. I won’t repeat the principles and concepts of RNA m6A today, and today I’m going to talk about the virus RNA m6A straightforwardly.
The impact of m6A modification on the life cycle of RNA viruses
It is known that epigenetic modification affects the life cycle of RNA viruses such as human coronaviruses. It has been reported that modified adenosine like N6-methyladenosine (m6A) affects the viability of specific RNA viruses by regulating virus cap structure, virus gene expression, virus progeny, innate sensing pathways, and innate immune response. The gain or loss of m6A can lead to significant functional changes of RNA viruses, changing host cell fusion/entry, replication, transmission, pathogen strength and immune evasion. The m6A phenotype of host cells plays an important role in host resistance, and it can also change after virus infection. The SARS-CoV-2 genome is more than 29,800 nucleotides in length. According to the presence of the specific sequence locus modified by the RNA methylase complex METL3/14 for m6A, including GGACU(T), GGACA and GGACC, it contains more than 50 potential The m6A site. Therefore, >0.64% of adenosine, or 0.18% of bases in SARS-CoV-2 RNA may be m6A. Therefore, the quantification of m6A in total RNA and the mapping of the whole transcriptome m6A map are essential for studying m6A-related viral functions and corresponding mechanisms, as well as identifying new antiviral therapeutic targets.
Two virus RNA extraction
Whether you start with a sample of cells, tissues or viruses, effective RNA isolation is usually the step to a successful experiment. For viruses: On the other hand, when it comes to virus samples, challenges such as low virus titer and high presence of contaminating host nucleic acid require the use of complementary techniques to ensure that yield and purity meet downstream requirements. In our most recent announcement article, we outlined 10 ways that this can be done, so you can apply it to the most suitable method for your study. One method is to inoculate cell cultures for viral amplification, and then collect the amplified virions for use with EpiGentek's viral RNA extraction kit. Note: For viral RNA extracted directly from clinical specimens, additional processing is required to remove co-extracted host DNA and rRNA contaminants before m6A RNA enrichment.
Three virus RNA extraction kit
There are two main types of viral RNA extraction products: RNA robust spin column and magnetic kits. Each of our kits is designed to extract high-quality RNA. In addition to RT-PCR, it can also be used Other downstream applications. Our kits differ in extraction methods and types of throughput options (high/manual), so no matter what your laboratory needs, our kits can help you get ready-to-use purified viral RNA. Our various magnetic beads and spin column kits can isolate RNA from different sample sources (nasal cavity/nasopharyngeal swabs, cells/tissues) and species (viruses, mammals). Detergents and deliquescent salts are used to lyse cells and inactivate RNase. The special high-salt buffer system allows RNA bases and magnetic beads to bind to the glass fiber matrix of the spin column, while contaminants are effectively washed away. Pure nucleic acid is eluted with water buffer, without phenol extraction or alcohol precipitation. The specific operation process is as follows:
P-9107 (rotating column) EpiQuik Viral RNA Isolation Fast Kit
EpiQuik Viral RNA Isolation Rapid Kit is a complete set of optimized buffers and reagents, suitable for rapid preparation of viral RNA from cell-free liquid samples (excluding plasma and serum), especially saliva and nasopharyngeal or nasopharyngeal swabs. Spin column purification. 10min.
P-9108 (magnetic bead method) EpiMag Viral RNA Isolation Kit (Magnetic Beads) is suitable for rapid preparation of viral RNA from cell-free liquid samples (excluding plasma and serum), especially saliva and nasopharyngeal or nasopharyngeal swabs. Bead method, 25min.
P-9109 (magnetic bead method) EpiMag 96-Well Viral RNA Extraction Kit (High Throughput) is suitable for preparing magnetic beads from cell-free liquid samples (without plasma and serum), and rapidly preparing viral RNA in a high-throughput manner. ), high flux, magnetic bead method, 30min.
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