High Pure Viral Nucleic Acid Large Volumen Kit
The High Pure Viral Nucleic Acid Large Volume Kit is designed for the purification of viral nucleic acids from up to 2.5 mL of mammalian serum, plasma or whole blood. When using whole blood, total nucleic acids are purified, including viral nucleic acids. The purified viral nucleic acids are applied in PCR or RT-PCR directly after elution in nuclease-free water.
Benefits of the High Pure Viral Nucleic Acid Large Volume Kit
- Improve sensitivity. Use sample volumes up to 2.5 ml to obtain high yields of purified nucleic acid in a concentrated 50 μl eluate.
- Obtain high-purity nucleic acids. Reduce carryover risk by using high centrifugal forces in all wash steps.
- Increase convenience and improve time to result. Eliminate complicated sample pre-processing and rapidly recover purified samples using high-speed centrifugation.
How the High Pure Viral Nucleic Acid Large Volume Kit works
As a pre-requisite for the analysis of viral nucleic acids by PCR or RT-PCR, the isolation of the analyte from serum, plasma, or whole blood is required. Virus lysis is accomplished by incubation of the sample in a special Lysis/Binding buffer in the presence of Proteinase K. Subsequently, nucleic acids bind specifically to the surface of glass fibers in the presence of a chaotropic salt. 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 surface of the glass fibers, thereby promoting adsorption to the glass fiber fleece. 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 eluted in low-salt buffer or water.
- Serum, plasma, or whole blood are lysed by incubation with Binding Buffer and Proteinase K.
- 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 get rid of PCR inhibitory contaminants.
- 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 HBV and HCV particles in human Citrate Plasma after isolation with the High Pure Viral Nucleic Acid Large Volume Kit and subsequent analysis on the LightCycler ® 480 Instrument II.
