High Pure Viral RNA Kit
The High Pure Viral RNA Kit efficiently purifies viral RNA from serum or plasma samples for direct RT-PCR analysis. This kit simplifies the purification process, eliminating the need for organic solvents or nucleic acid precipitation, and facilitates the rapid preparation of RT-PCR templates. Utilizing a special Binding Buffer and glass fibers, nucleic acids are specifically bound and purified, while an Inhibitor Removal Buffer enhances RT-PCR assay sensitivity by removing contaminants. This streamlined process ensures the extraction of high-quality viral RNA, suitable for accurate RT-PCR analysis, in just minutes.
Benefits of the High Pure Viral RNA Kit
- Saves time by avoiding use of organic solutions or nucleic acid precipitation; prepare multiple RT-PCR templates in approximately 10 minutes.
- Purify viral RNA from a variety of samples.
- Minimize RNA loss with a kit that removes contaminants without precipitation or solvent extraction.
- Increase lab safety by minimizing the handling of potentially hazardous samples avoiding hazardous organic solvents.
How the High Pure Viral RNA Kit works
As a pre-requisite for the analysis of viral RNA by the reverse transcription polymerase chain reaction (RT-PCR), the isolation of the analyte from serum or plasma is required. The High Pure Viral RNA Kit accomplishes virus lysis by incubation of the sample in a special Binding Buffer. Subsequently, nucleic acids bind specifically to the surface of glass fibers in the presence of a chaotropic salt (guanidine-HCl). The binding reaction occurs within seconds due to the disruption of the organized structure of water molecules and the interaction of nucleic acids with the glass fibers surface. Thus, adsorption to the glass fiber fleece is favored.
Since the binding process is specific for nucleic acids, the bound nucleic acids are purified from salts, proteins, and other impurities by a washing step and are finally eluted in low-salt Elution Buffer or PCR-grade water. The purified viral RNA is free of intact virus, nucleases, and all cellular components that interfere with RT-PCR, and can be applied directly for RT-PCR. Fifty microliter eluate is sufficient for 8 to 14 RT-PCR reactions.
Included in the kit is a special Inhibitor Removal Buffer that results in improved sensitivity and reproducibility of RT-PCR assays performed with nucleic acid templates isolated with this kit. The use of the Inhibitor Removal Buffer allows even the application of heparinized sample material containing 100 U/mL heparin.
- Serum or plasma are lysed by incubation with Binding Buffer.
- Nucleic acids are bound to the glass fibers pre-packed in the High Pure Filter Tube.
- Bound nucleic acids are washed with a special Inhibitor Removal Buffer to remove RT-PCR inhibitory contaminants. – Allows even the application of heparinized sample material with >100 U/mL heparin.
- Washing of bound nucleic acids, purification from salts, proteins. and other cellular impurities.
- Purified nucleic acids are recovered using the Elution Buffer.
Figure: Crossing Points of a series dilution of HCV particles in human Citrate Plasma after isolation with the High Pure
Viral RNA Kit and subsequent analysis on the LightCycler® 480 Instrument II.
