High Pure Viral RNA Kit
Low to medium throughput viral RNA isolation.
For general laboratory use.
The High Pure Viral RNA Kit is for general laboratory use and purifies viral RNA from a variety of samples such as serum and plasma.
- Remove inhibitors that interfere with downstream assays, ensuring greater assay specificity, sensitivity, and reproducibility.
- Prepare RNA samples in only 20 minutes.
- Obtain concentrated RNA that is suitable for downstream applications.
- Minimize RNA loss with a kit that removes contaminants without precipitation or other handling steps that degrade RNA.
- Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.
- Binding Buffer
- Poly (A), lyophilizate
- Inhibitor Removal Buffer
- Wash Buffer
- Elution Buffer
- High Pure Spin Filter Tubes
- Collection Tubes
The High Pure Viral RNA Kit rapidly isolates viral RNA from mammalian plasma, serum, body fluids, and cell culture supernatants. The isolated RNA is suitable for RT-PCR after elution in nuclease-free water (see Figure 1).
Figure 1: RT-PCR analysis of MS2 RNA isolated with the High Pure Viral RNA Kit.
Serial dilutions of purified MS2 RNA were applied to High Pure filter tubes, processed according to the kit protocol, and eluted in a total volume of 50 µL. An aliquot (3.5 µL) of each eluate was analyzed by two-step RT-PCR, using primers that produced a 961 bp product. Products were analyzed by agarose gel electrophoresis. For each sample listed below, the "number of molecules per PCR" assumes that all the RNA applied to the High Pure tube was recovered in the eluate.
Lane 1: 3.5 x 107 molecules/RT-PCR
Lane 2: 3.5 x 105 molecules/RT-PCR
Lane 3: 3.5 x 103 molecules/RT-PCR
Lane 4: 3.5 x 102 molecules/RT-PCR
Lane 5: 3.5 x 101 molecules/RT-PCR
Lane M: DNA Molecular Weight Marker III
Result: Even when the template contained a theoretical maximum of 35 molecules viral RNA, RT-PCR generated a product that was detectable on an agarose gel.
Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. For RNA isolation, the binding conditions can be optimized to ensure immobilization of all the RNA.
Note: The isolation process includes the addition of a special Inhibitor Removal Buffer. This buffer increases the sensitivity and reproducibility of RT-PCRs performed with the isolated viral RNA, even when the sample contains heparin.
Capacity: The High Pure Spin Filter Tubes hold up to 700 μl sample volume.
Typical Nucleic Acid Recovery
Starting Material and Quantity
Number of Reactions
|Serum, plasma, urine, supernatant from cell culture, 200 - 600 µL||Product detectable by RT-PCR||10 minutes||100/200 µL|
Viruses, when lysed by a detergent and Proteinase K, release total viral nucleic acids. In the presence of a chaotropic salt (guanidine HCl), viral nucleic acids bind selectively to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, viral RNA binds to the glass fiber fleece, while contaminating substances (salts, proteins, and other cellular contaminants) do not. Brief wash-and-spin steps readily remove the contaminants. The remaining, purified RNA is then eluted in a small volume of low-salt buffer.
Series of MS2 RNA dilutions are prepared, applied to the filter tubes, washed, and eluted following the kit protocol. 3.5 μL of the eluate is analyzed by RT-PCR. The products are detected on agarose gel. At least 2 x 105 RNA molecules/200 μL sample are guaranteed.