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High Pure Viral RNA Kit Product No. Pack Size 11858882001 100 purifications Roche
High Pure Viral RNA Kit
Low to medium throughput viral RNA isolation.
For general laboratory use.
| Product No. | Pack Size |
|---|---|
| 11858882001 | 100 purifications |
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.
Contents
- 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 |
Yield/Recovery |
Time Required |
Number of Reactions |
| Serum, plasma, urine, supernatant from cell culture, 200 - 600 µL | Product detectable by RT-PCR | 10 minutes | 100/200 µL |
Principle
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.
Quality
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.
Alternate DE MSDS - Material Safety Data Sheet
Alternate DE Instructions for Use - Version 19
Select the Version Number printed on the outer label of the product box.
To select documents, click on the download button shown beside.
Nucleic Acid Isolation and Purification Product Selection Guide
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