For viral RNA isolation from cell-free body fluids

Features

Rapid isolation of high-quality, ready-to-use RNA

No organic extraction or alcohol precipitation

Complete removal of contaminants and inhibitors

RNA isolation from viruses, such as SARS-CoV-2, up to 96-well format

Automated purification from low to ultra-high throughput format

Product Details

QIAamp Viral RNA Kits simplify viral RNA purification from cell-free body fluids with fast spin-column, vacuum and plate centrifugation 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 the QIAcube Connect using the QIAamp Viral RNA Mini Kit or dedicated QIAamp Viral RNA Mini QIAcube Kit. Various options for automation of the QIAamp 96 Viral Kit on instruments including the QIAcube HT, the Fluent 780 from Tecan or the epMotion 5075V from Eppendorf support high to ultra-high throughput processing with minimal hands-on time.

QIAamp Viral RNA Mini and QIAamp 96 Viral RNA standard protocols can also be executed using the TRACKMAN Connected system, paired with PIPETMAN M Connected pipettes, both from Gilson. The TRACKMAN Connected system guides researchers through QIAamp Viral RNA Mini or QIAamp 96 Viral RNA protocols while automatically adjusting the Bluetooth-enabled PIPETMAN M Connected pipette settings. Each step of the protocol execution is recorded to accelerate reporting by generating a comprehensive run report.

Performance

The high-quality viral RNA isolated using the QIAamp Viral RNA Mini Kit and QIAamp Viral RNA Mini QIAcube Kit performs well in a wide range of downstream applications, including viral genotyping, viral epidemiology and infectious disease research.

The QIAamp Viral RNA Mini QIAcube Kit includes rotor adapters that are preloaded with QIAamp spin columns and elution tubes, delivering greater convenience and time savings . Furthermore, ease of use is increased and user errors minimized. Waste is reduced because the content of the dedicated kit is tailored for purification on the QIAcube Connect and the superfluous tubes that are required for the manual procedure are not included.

The QIAamp 96 Viral RNA Kit has been successfully used to recover RNA from viruses, including SARS-CoV-2 virus . Performance of the QIAamp 96 Viral RNA Kit is comparable to that achieved with the QIAamp Viral RNA Mini Kit . RNA isolation is proven from commonly used saliva collection and stabilization methods and no cross-contamination was detected .

Principle

The QIAamp Viral RNA Mini Kit and QIAamp Viral RNA Mini QIAcube Kit simplify viral RNA purification from cell-free body fluids with fast spin-column, vacuum or automated procedures. The QIAamp 96 Viral RNA Kit simplifies viral RNA isolation from nasotracheal swabs. 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 or 96-well plate. Wash buffers are used to remove impurities and pure, ready-to-use RNA is then eluted in water or low-salt buffer.

Vacuum processing

For greater speed and convenience in RNA purification, samples can be processed by vacuum instead of centrifugation. QIAamp Mini spin columns 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 to ensure consistent vacuum. Disposable VacConnectors are used to avoid any cross-contamination.

Automated processing on the QIAcube Connect

A dedicated QIAamp Viral RNA QIAcube Kit enables automated processing on the QIAcube Connect. In addition, the QIAamp Viral RNA Mini Kit is automatable on the QIAcube Connect. The QIAamp Viral RNA Mini Accessory Set provides the additional buffers and reagents needed for automated, low-throughput sample prep using both the QIAcube Connect and QIAamp Viral RNA Mini Kit.

Processing in 96-well format

The QIAamp 96 Viral RNA Kit is automatable on various instruments.

We recommend the use of the Centrifuge 4-16S or Centrifuge 4-16KS for centrifuge processing. Alternative centrifuges with swing-out rotor, minimum 5788 x g capacity and the ability to accommodate plate assemblies with a height of minimum 75 mm or more may be used. Centrifuges with lower g-force (minimum 3486 x g) can be used with an alternative protocol.

Plasticware recommended for centrifugation processing: Plastic consumables need to be in SBS format and must be able to withstand the necessary g-forces. Deep-well plates need to have a capacity of 2 ml and elution plates need to have a capacity of ≥500 µl. Four deep-well plates and one elution plate are required for processing of 96 samples.

Applications

QIAamp Viral RNA Kits simplify viral RNA purification from cell-free body fluids, offering fast spin-column, vacuum or plate centrifugation procedures, or automation on QIAcube. Sample types include:

Plasma and serum

CSF

Urine

Other cell-free body fluids

Cell-culture supernatants

Swabs

Supporting data and figures

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.