QIAamp MinElute Virus Kits

For simultaneous purification of viral DNA and RNA from plasma, serum and cell-free body fluids

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QIAamp MinElute Virus Spin Kit (50)

Cat. No. / ID: 57704

For 50 minipreps: 50 QIAamp MinElute Columns, QIAGEN Protease, carrier RNA, buffers, Collection Tubes (2 ml)

KitAccessories

QIAamp MinElute Virus Kit

QIAamp MinElute Virus Accessory Set

Carrier RNA

For

Spin column procedure

Vacuum procedure

Inquire

QIAamp MinElute Virus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Maximal concentration in eluate for high downstream sensitivity
Rapid purification of high-quality viral DNA and RNA
No organic extraction or alcohol precipitation
Consistent, high yields
Removal of contaminants and inhibitors

Product Details

QIAamp MinElute Virus Kits simplify viral DNA and RNA purification with fast spin-column and vacuum procedures. The kits use starting sample volumes of up to 0.2 ml and combine the selective binding properties of a silica-based membrane with flexible elution volumes of between 20 and 150 μl. The QIAamp MinElute Virus Spin process can be fully automated on the QIAcube Connect. The additional buffers and reagents in the QIAamp MinElute Virus Accessory are required for automation of the QIAamp MinElute Virus Spin procedure.

Performance

Purified viral nucleic acids are free of proteins, nucleases and other impurities, and are suitable for use in sensitive downstream applications such as PCR and RT-PCR (see figures " High sensitivity in PCR" and " High sensitivity in RT-PCR").

Viral nucleic acids purified using QIAamp MinElute Virus Kits can be used in a wide range of downstream applications, including:

PCR and quantitative real-time PCR
Infectious disease research

See figures

High sensitivity in PCR.

High sensitivity in RT-PCR.

Principle

QIAamp MinElute Virus Kits simplify the isolation of viral RNA and DNA from plasma, serum and cell-free body fluids with a fast spin-column or vacuum procedure. No phenol–chloroform extraction is required. Nucleic acids bind specifically to the QIAamp MinElute silica-gel membrane while contaminants pass through. PCR inhibitors, such as divalent cations and proteins, are completely removed in two efficient wash steps, leaving pure nucleic acids to be eluted in either water or a buffer provided with the kit. QIAamp MinElute technology yields viral DNA and RNA from serum, plasma and cell-free body fluids that are ready to use in PCR and blotting procedures.

QIAamp MinElute Virus Kits combine the selective binding properties of a silica-based membrane with flexible elution volumes of between 20 and 150 µl. The QIAamp MinElute Virus Spin procedure can be fully automated on the QIAcube Connect. The additional buffers and reagents provided in the QIAamp MinElute Virus Accessory Set are required.

QIAamp sample preparation technology is fully licensed.

Procedure

Optimized buffers lyse samples, stabilize nucleic acids and enhance selective DNA adsorption to the QIAamp MinElute membrane (see flowcharts " QIAamp MinElute Virus Spin procedure" and “ QIAamp MinElute Virus Vacuum procedure”). Alcohol is added and lysates loaded onto the QIAamp MinElute spin column. Wash buffers are used to remove impurities and viral nucleic acids are eluted in Buffer AVE, ready for use in amplification reactions or storage at –20ºC. Purified nucleic acids are free of proteins, nucleases and other impurities.

See figures

QIAamp MinElute Virus Spin procedure.

QIAamp MinElute Virus Vacuum procedure.

Applications

The QIAamp MinElute Virus Spin Kit uses well-established technology for simultaneous purification of viral RNA and DNA from fresh or frozen plasma, serum, other cell-free body fluids.

The QIAamp MinElute Virus Vacuum Kit uses well-established technology for simultaneous purification of viral RNA and DNA. Fresh or frozen plasma, serum and other cell-free body fluids (500 µl) can be processed in less than an hour.

Comparison of QIAamp MinElute Virus Kits

Features	QIAamp MinElute Virus Spin Kit	QIAamp MinElute Virus Vacuum Kit
Applications	PCR, real-time PCR	PCR, real-time PCR
Elution volume	20–150 µl	20–150 µl
Format	MinElute columns	MinElute columns
Main sample type	Serum, plasma	Serum, plasma
Processing	Manual (centrifugation)	Manual (vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Viral DNA, viral RNA	Viral DNA, viral RNA
Sample amount	200 µl	500 µl
Technology	Silica technology	Silica technology
Time per run or per prep	<1 hour	<1 hour
Yield	Varies	Varies

Supporting data and figures

High sensitivity in PCR.

The percentage of positive samples at low virus titers is shown. Nucleic acids from a DNA virus were purified from plasma using either the QIAamp MinElute Spin or Vacuum Kit and subjected to PCR. For the spin procedure, 95% probit values for 40 samples were 27.25 IU/ml and for the vacuum procedure, they were 9.30 IU/ml.

Resources

For simultaneous purification of viral RNA and DNA from plasma, serum, and cell-free body fluids

This procedure has been adapted by customers and is for purification of viral RNA and DNA from plasma, serum, and cell-free body fluids using the QIAamp MinElute Virus Vacuum Kit.

Publications

Verification of an assay for quantification of hepatitis C virus RNA by use of an analyte-specific reagent and two different extraction methods.

Forman MS; Valsamakis A;

J Clin Microbiol; 2004; 42 (8):3581-8 2004 Aug PMID:15297501

Real-time PCR assay for detection and quantification of hepatitis B virus genotypes A to G.

Welzel TM; Miley WJ; Parks TL; Goedert JJ; Whitby D; Ortiz-Conde BA;

J Clin Microbiol; 2006; 44 (9):3325-33 2006 Sep PMID:16954268

Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing.

Breitbart M; Rohwer F;

Biotechniques; 2005; 39 (5):729-36 2005 Nov PMID:16312220

Prevalence and stability of human serum antibodies to simian virus 40 VP1 virus-like particles.

Lundstig A; Eliasson L; Lehtinen M; Sasnauskas K; Koskela P; Dillner J;

J Gen Virol; 2005; 86 (Pt 6):1703-1708 2005 Jun PMID:15914848

FAQ

Yes, Carrier RNA is available separately. It is sold in combination with other buffers in the QIAamp MinElute Virus Accessory Set and the QIAamp Viral RNA Mini Accessory Set.

In addition, Poly-A Carrier RNA by itself, without any buffers, is offered under the following catalog numbers:

•1017647: 12 vials, each containing 1350 µg lyophilized carrier RNA

•1068337: 1 vial containing 310 µg lyophilized carrier RNA

FAQ ID -351

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560

Aside from the obvious difference of using a centrifuge versus a vacuum manifold for sample processing, the QIAamp MinElute Virus Spin Kit requires sample volumes of 200 ul, whereas the QIAamp MinElute Virus Vacuum Kit can process 500 ul of sample due to Extension tubes provided in the kit.

FAQ ID -295

Yes, it should be possible to use the QIAamp MinElute Virus Kits for this purpose. Several customers have successfully isolated viral nucleic acid from cell culture supernatant by following the standard protocols of the QIAamp MinElute Virus Spin Handbook or the QIAamp MinElute Virus Vacuum Handbook.

FAQ ID -472

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

FAQ ID -761

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.

For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.

FAQ ID -635

For RNA isolated on the BioRobot EZ1 and BioRobot M48:

The RNA can be directly applied to the Agilent Bioanalyzer, since it is being denatured in the final protocol steps of these isolation procedures.

For RNA prepared with all other QIAGEN RNA Isolation Products:

We recommend to denature the samples in a water bath for 2 min at 70°C, and then place them directly on ice prior to loading them onto the Agilent Bioanalyzer.

FAQ ID -528

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12