For isolation of viral RNA from cell-free body fluids
Rapid isolation of high-quality, ready-to-use RNA
No organic extraction or alcohol precipitation
Consistent, high yields
Complete removal of contaminants and inhibitors
The QIAamp Viral RNA Mini Kit simplifies purification of viral RNA from cell-free body fluids with fast spin-column or vacuum procedures. Viral RNA binds specifically to the QIAamp silica membrane, and pure viral RNA is eluted in either water or a buffer provided with the kit. Purification can be fully automated on theQIAcube Connect.
For 250 RNA preps: 250 QIAamp Mini Spin Columns, carrier RNA, Collection Tubes (2 ml), RNase-free buffers
The QIAamp Viral RNA Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Amplification of RNA from plasma.
RT-PCR products of a 1026 nt RNA fragment purified from plasma. Serial tenfold dilutions (as indicated) were added to plasma and purified using the QIAamp Viral RNA Mini Kit. M: markers; C: negative control.
Performance
The high-quality viral RNA isolated using the QIAamp Viral RNA Mini Kit performs well in a wide range of downstream applications, including viral genotyping, viral epidemiology, and infectious disease research (see figure "Amplification of RNA from plasma").
Principle
The QIAamp Viral RNA Mini Kit simplifies isolation of viral RNA from cell-free body fluids with fast spin-column or vacuum procedures. No phenol–chloroform extraction is required. Viral RNA binds specifically to the QIAamp silica membrane while contaminants pass through. PCR inhibitors, such as divalent cations and proteins, are completely removed in two efficient wash steps, leaving pure viral RNA to be eluted in either water or a buffer provided with the kit.
QIAamp RNA technology yields viral RNA from cell-free body fluids ready to use in RT-PCR and blotting procedures. QIAamp sample preparation technology is fully licensed.
Procedure
Optimized buffers and enzymes lyse samples, stabilize nucleic acids, and enhance selective RNA adsorption to the QIAamp membrane. To guarantee RNA integrity, samples are lysed under highly denaturing conditions to inactivate RNases. Alcohol is added and lysates loaded onto the QIAamp spin column. Wash buffers are used to remove impurities and pure, ready-to-use RNA is then eluted in water or low-salt buffer.
The QIAamp Viral RNA Mini Kit is automatable on the QIAcube. The QIAamp Viral RNA Mini Accessory Set provides the additional buffers and reagents needed for automated, low-throughput sample prep using both QIAcube and QIAamp Viral RNA Mini Kit.
Vacuum processing
For greater speed and convenience in RNA purification, samples can be processed by vacuum instead of centrifugation. QIAamp Mini spin columns (see figure "QIAamp Viral RNA Mini spin column") are accommodated on the QIAvac 24 manifold using VacValves and VacConnectors, provided in the QIAamp Vac Accessory Set. VacValves should be used if sample flow rates differ significantly, in order to ensure consistent vacuum. Disposable VacConnectors are used to avoid any cross-contamination.
Applications
The QIAamp Viral RNA Mini Kit simplifies purification of viral RNA from cell-free body fluids, offering fast spin-column or vacuum procedures, or automation on QIAcube. Sample types include:
Plasma and serum
CSF
Urine
Other cell-free body fluids
Cell-culture supernatants
Features
Specifications
Applications
PCR, qPCR, real-time PCR
Elution volume
50 µl
Format
Spin columns
Main sample type
Liquid media
Processing
Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
For 100 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 200 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 1 ml), dNTP Mix (1 x 200 µl, 10 mM each), 5x Q-Solution (1 x 2 ml), RNase-Free Water (2 x 1.9 ml)
For 100 x 20 µl reactions: 1 x 500 µl QuantiNova Pathogen Master Mix, 1 x 500 µl Yellow Template Dilution Buffer, 1 x 250 µl ROX Reference Dye, 1 x 1.9 µl RNase-Free Water, 1 x 1500 µl Nucleic Acid Dilution Buffer, 1 x 100 µl Internal Control RNA, 1 x 100 µl Internal Control DNA, 1 x 200 µl IC Probe Assay
Bag of 50; suitable for holding diluent and master mix on the instrument worktable; graduated, flat-base design with a tapered internal profile for minimum dead volume; screw cap included