QIAamp Virus BioRobot MDx Kit

For automated purification of viral DNA and RNA from cell-free body fluids on the BioRobot MDx workstation

Products

The QIAamp Virus BioRobot MDx Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

QIAamp Virus BioRobot MDx Kit (12)

Cat. No. / ID: 965652

For 12 x 96 preps for use on the BioRobot MDx; includes: 12 QIAamp 96 Plates, RNase-free buffers, QIAGEN Protease, Elution Microtubes CL, caps, S-Blocks, carrier RNA

47.296,00 BRL

Quantity

Features

Rapid isolation of high-quality, ready-to-use viral DNA and RNA
No organic extraction or alcohol precipitation
Consistent, high yields
Removal of contaminants and inhibitors

Product Details

The QIAamp Virus BioRobot MDx Kit provides automated viral nucleic acid purification on the BioRobot MDx using proven QIAamp technology. The fully automated procedure requires less than 2.5 hours, including bar code reading and complete process documentation, with no hands-on time during the run.

Performance

The QIAamp Virus BioRobot MDx process provides uniform DNA and RNA recovery and purity across the 96-well plate with no detectable cross-contamination between samples. Up to 265 µl of cell-free body fluids can be processed. The typical elution volume is 75 µl. Viruses can be detected with high sensitivity when using this kit, which is illustrated by hit-rate studies (see table). Further advantages of the system include high process reliability and walkaway automation, providing a highly efficient system for purification of viral nucleic acids. The QIAamp Virus BioRobot MDx provides reliable results in downstream applications.

High precision using the QIAamp Virus BioRobot MDx Kit
	Mean C_T	Standard deviation	%CV	Mean C_T	Standard deviation	%CV
PCR product (n=96)	Instrument 1			Instrument 2
Mimic virus	28.26	0.10	0.35	28.38	0.18	0.65
Internal control	31.98	0.16	0.51	31.87	0.22	0.70

Plasma was spiked with armored RNA molecules to mimic RNA viruses (25,000 copies per prep) and processed using the QIAamp Virus BioRobot MDx protocol in replicates of 96 on two separate instruments. An internal control RNA was added to the lysis buffer. The eluted RNA was amplified using a TaqMan® RT-PCR system. Mean C_T value, standard deviation, and %CV were calculated for both the mimic virus and the internal control. (Data generated by QIAGEN R&D department.)

Principle

The QIAamp Virus BioRobot MDx Kit simplifies isolation of viral RNA and DNA from cell-free body fluids with a fast automated 96-well–plate procedure. No phenol–chloroform extraction is required. Nucleic acids bind specifically to the QIAamp 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 technology yields viral RNA and DNA from cell-free body fluids ready to use in PCR and blotting procedures.

The fully automated procedure, including bar code reading and complete process documentation, requires less than 2.5 hours to process 96 samples (including a detailed load check of 20 minutes) with no hands-on time during the run. Advantages of the system include high process reliability and walkaway automation, providing a highly efficient system for purification of viral nucleic acids for laboratories. QIAamp sample preparation technology is fully licensed.

Procedure

Optimized buffers lyse samples, stabilize nucleic acids, and enhance selective nucleic acid adsorption to the QIAamp membrane. Alcohol is added and lysates loaded onto the QIAamp 96-well plate. Wash buffers are used to remove impurities and pure, ready-to-use nucleic acids are then eluted in water or low-salt buffer (see flowchart "").

See figures

QIAamp Virus BioRobot MDx protocol.

Applications

The QIAamp Virus BioRobot MDx Kit provides automated, high-throughput purification of viral RNA and DNA on the BioRobot MDx using proven QIAamp technology. The 96-well plates provide the flexibility of processing 32–96 samples, in increments of 8. Sample sources include:

Plasma and serum
CSF
Other cell-free body fluids
Cell-culture supernatants

Supporting data and figures

QIAamp Virus BioRobot MDx protocol.

Specifications

Features	Specifications
Applications	PCR, qPCR, real-time PCR
Samples per run (throughput)	36-92 samples per run
Technology	Silica technology
Main sample type	Serum, plasma
Elution volume	75 µl
Processing	Automated
For automated processing	BioRobot MDx Workstation
Sample amount	265 µl
Format	96-well plates
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Viral DNA, viral RNA
Time per run or per prep	<2.5 hours
Yield	>90% recovery

Resources

For simultaneous purification of viral RNA and DNA from plasma, serum, cell-culture supernatants, and cell–free body fluids using the BioRobot MDx

FAQ

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

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