Description

For high-throughput purification of viral RNA
  • Suitable for RNA isolation from viruses, such as SARS-CoV-2
  • Consistent and reliable performance in 96-well format
  • Choice of manual or automated processing
  • Compatible with various plastic consumables and instruments for automation
  • Efficient removal of inhibitors and contaminants
  • Purified nucleic acids ready for analysis by real-time RT-PCR

The QIAamp 96 Viral RNA Kit enables simple, purification of viral RNA manually with a plate centrifuge. Various options for automation on instruments including the Fluent 780 from Tecan, or the epMotion 5075V from Eppendorf support high-throughput processing with minimal hands-on time. Using proven QIAamp silica-membrane technology in a convenient 96-well format, contaminants and inhibitors are removed to yield high-quality nucleic acids directly ready for downstream analysis.

Performance

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

Principle

The QIAamp 96 Viral RNA Kit simplifies the isolation of viral RNA 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.

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 96 Plate. Wash buffers are used to remove impurities and pure, ready-to-use RNA is then eluted in water or low-salt buffer. 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 the 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 the processing of 96 samples.

Applications

The QIAamp 96 Viral RNA Kit simplifies high-throughput viral RNA, offering fast 96-well procedures, or automation on suitable centrifuges and instruments.

Specifications

Features
Specifications
Elution volume80 µl
Number of preps per run24–96 samples (processed in increments of 8)
Sample size140 µl solution from a swab sample
Throughput96 samples in approximately 90 minutes with centrifuge Up to 4 x 96 samples in approximately 60 minutes with sample processing instruments