Viral Rna Isolation
Viral Rna Isolation - Related Videos
18:10
Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
0 Views •
Cited by 40 •
2011
The present article describes the steps required to isolate and characterize RNA polymerase fidelity variants of RNA viruses and how to use mutation frequency data to confirm fidelity changes in tissue culture.
10:16
Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
0 Views •
Cited by 35 •
2012
A fluorescence in situ hybridization (FISH) method was developed to visually detect viral genomic RNA using fluorescence microscopy. A probe is made with specificity to the viral RNA that can then be identified using a combination of hybridization and immunofluorescence techniques. This technique offers the advantage of identifying the localization of the viral RNA or DNA at steady-state, providing information on the control of intracellular virus trafficking events.
02:59
Extraction of Viral RNA from Surface-Bound Virions Using a Silica Spin Column
0 Views •
2026
Source: Klaus, J. P. et al. Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment. J. Vis. Exp. (2016)This video demonstrates the extraction of viral RNA from surface-bound virions on epithelial cells using silica spin columns. The method includes chemical lysis, RNA stabilization, and membrane-based purification for downstream quantification of virus-cell attachment.
09:35
Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis
0 Views •
Cited by 15 •
2013
This method describes the use of click chemistry to measure changes in host cell transcription after infection with the Rift Valley fever virus (RVFV) strain MP-12. Results can be visualized qualitatively via fluorescence microscopy or obtained quantitatively through flow cytometry. This method is adaptable for use with other viruses.
09:43
Isolation and Characterization of RNA-Containing Exosomes
0 Views •
Cited by 515 •
2012
This paper demonstrates methods for the isolation, purification and detection of exosomes, as well as techniques for analysis of their molecular content. These methods are adaptable for exosome isolation from both cell culture media and biological fluids, and can beyond analysis of molecular content also be useful in functional studies.
FAQs
Chaotropic reagents serve two linked purposes: they help disrupt viral particles and inactivate contaminating ribonucleases, enzymes that can degrade RNA. This creates conditions in which viral RNA remains available for capture rather than being destroyed during sample processing. Effective inactivation is therefore central to obtaining material suitable for sensitive downstream molecular analyses.
Silica membranes and magnetic beads provide solid surfaces that capture RNA after viral particles have been disrupted and ribonucleases have been inactivated. Washing removes unwanted components while the captured RNA remains associated with the solid phase, and elution releases it for later analysis. These materials form the purification stage of the workflow.
RNase-free handling helps prevent degradation of the target molecule before and during purification. Because the objective is to preserve intact viral RNA, contamination with ribonucleases can reduce the amount and quality of material recovered. Maintaining careful conditions improves the reliability of downstream assays and supports more consistent comparisons among biological samples.
A typical workflow begins with a biological sample and disruption of viral particles. Chaotropic reagents then inactivate contaminating ribonucleases, after which RNA is captured on a silica membrane or magnetic beads. The solid phase is washed to remove unwanted material, and the purified RNA is finally eluted for downstream testing or characterization.
The integrity and cleanliness of recovered viral RNA influence how reliably it can support reverse transcription polymerase chain reaction, sequencing, and viral detection. Well-preserved material can improve assay sensitivity and provide a stronger basis for genome characterization. Poor preservation may limit the information available for comparing viral variants or studying infection-related patterns.
Researchers can apply the purified material to detect viruses, characterize viral genomes, compare variants, and investigate infection and transmission. The resulting analyses also support outbreak monitoring and studies of host-pathogen interactions. Thus, the technique connects sample processing with both immediate detection goals and broader investigations of viral biology.