Viral RNA isolation is similar to non-viral RNA extraction or RNA purification
Viral RNA Isolation: what is it and how does it help my studies?
RNA isolation is the process by which RNA is separated from viruses, cells, or sample matrices. RNA (ribonucleic acid) is a polymeric molecule essential in various biological roles in coding, decoding, regulation and expression of genes. RNA is often extracted for the purpose of downstream PCR applications.
You can learn more about RNA isolation HERE
You can learn more about RNA isolation HERE
Viral RNA can be isolated directly from serum, plasma, culture medium, or other cell-free fluids by means of specially formulated spin columns or beads. Cell-free material is used for these isolations because most total RNA isolation methods cannot distinguish between RNA from microorganisms and those from host cells. If cell-free material is not readily available, concentration of viroids via centrifugation is also used.
Isolating the genetic material of a virus has proven to contribute greatly to medicine and molecular diagnostics. Once the RNA is isolated, amplification methods such as polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) can be used for detection of viral pathogens such as HIV, coronaviruses and others. Because of RNase activity is a primary concern for non-viral RNA isolations, it becomes even more dangerous in the delicate isolation of viral RNA. In response to this, manufacturers make the extraction buffers sterile and RNase free (RNF). The delicate viral RNA is most optimally used when the assay will be carried out immediately, for instance in infectious disease testing (i.e., COVID-19).
Isolating the genetic material of a virus has proven to contribute greatly to medicine and molecular diagnostics. Once the RNA is isolated, amplification methods such as polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) can be used for detection of viral pathogens such as HIV, coronaviruses and others. Because of RNase activity is a primary concern for non-viral RNA isolations, it becomes even more dangerous in the delicate isolation of viral RNA. In response to this, manufacturers make the extraction buffers sterile and RNase free (RNF). The delicate viral RNA is most optimally used when the assay will be carried out immediately, for instance in infectious disease testing (i.e., COVID-19).
How is RNA isolated?
Solid-phase viral RNA isolation uses beads or columns to capture and release RNA at desired times. The initial procedure is similar to organic isolation, where a lysate is first obtained. This sample is incubated with binding beads or columns, that selectively bind to RNA and thereby allows for the removal of proteins, salts, PCR inhibitors, and other contaminants. Magnetic beads or columns are often used in automated systems and are available in convenient Viral RNA isolation kits.
- Magnetic beads selectively bind to RNA in the lysate and are then separated from the lysate using a magnet. These beads are washed to remove contaminants, before the RNA is eluted off of the beads, resulting in isolated RNA.
- Columns selectively bind to RNA in the lysate and allow non-RNA components of the lysate to flow through the column. The column is then washed to remove contaminants, before the RNA is eluted off of the column, resulting in isolated RNA.
Direct PCR / extraction-free PCR and the improved, Next Generation Direct PCR, allows for downstream PCR applications without the need for viral RNA extraction / viral RNA purification. Furthermore, direct PCR does not alter the RNA profile of your sample since it does not use 'capture and release' technology or separate components of your sample with organic solvents like Trizol, as used by solid-phase and organic RNA isolation.
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