QIAamp System

The QIAamp system is designed to purify genomic, mitochondrial, and bacterial DNA, total cellular RNA, or viral nucleic acids from a wide range of clinical samples for downstream amplification and blotting applications. QIAamp sample preparation technology is fully licensed, allowing QIAamp purified nucleic acids to be used in any molecular assay or other downstream application without risk of patent infringement.

Features and benefits

• Rapid isolation of high-quality, ready-to-use DNA or RNA
• No organic extraction or alcohol precipitation
• Consistent, high yields
• Complete removal of contaminants and inhibitors for reliable downstream amplification, blotting, or restriction digestion
• Vacuum-driven processing for some QIAamp Kits, for more convenient purification*

Principle

QIAamp Kits simplify isolation of nucleic acids from a wide variety of clinical samples with fast spin-column or 96-well–plate procedures. 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 acid to be eluted in either water or a buffer provided with the kit. QIAamp technology yields genomic, mitochondrial, and bacterial DNA, total cellular RNA, and viral nucleic acids from many sample sources ready to use in PCR and blotting procedures.

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 spin column or 96-well plate. Wash buffers are used to remove impurities and pure, ready-to-use DNA or RNA is then eluted in water or low-salt buffer. The entire process requires only 20 minutes of handling time for mini spin-column formats (lysis times differ according to the sample source). Processing time using high-throughput QIAamp 96 Kits is as little as 1 minute per sample.

Vacuum processing

With the QIAamp DNA Blood Mini Kit, the QIAamp DNA Mini Kit, and the QIAamp Viral RNA Mini Kit, blood and other body fluids can be processed by vacuum instead of centrifugation, for greater speed and convenience in nucleic acid purification. 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, in order to ensure consistent vacuum. Disposable VacConnectors are used to avoid any cross-contamination. Use of VacConnectors also allows these QIAamp spin procedures to be performed on QIAvac 6S with QIAvac Luer Adapters.

Downstream applications

DNA purified using QIAamp DNA Kits is sized up to 50 kb (see figure "Purified Genomic DNA up to 50 kb"). DNA of this length denatures completely and has the highest amplification efficiency. RNA purified with QIAamp RNA Kits is ideal for use in applications such as RT-PCR and northern blotting. QIAamp Kits are validated for general-purpose use in a number of diagnostic applications,* including:

• Viral diagnostics
• Bacterial diagnostics
• Fungal diagnostics
• HLA typing (see figure "HLA Typing by Nested SSP-PCR")
• Cancer research
• Human genetic testing
• Paternity testing (see figure "Paternity Testing by RFLP Analysis")
• Forensic analysis

* QIAamp Kits are intended as general-purpose devices. No claim or representation is intended for their use to identify any specific organism or for clinical use (diagnostic, prognostic, therapeutic, or blood banking). It is the user’s responsibility to validate the performance of QIAamp Kits for any particular use, since the performance characteristics of these kits have not been validated for any specific organism.

Purified Genomic DNA up to 50 kb

Size distribution of DNA prepared with QIAamp Kits from the indicated sources (3 µg per lane).

HLA Typing by Nested SSP-PCR

PCR products obtained with QIAamp purified DNA from a dialysis patient and HLA-DRB1–specific primers. D: HLA-DRB1 1610/1602 (DR16), 144 bp fragment. L: HLA-DRB1 1101–1105 (DR11), 130 bp fragment. M1, M2: markers. (Data kindly provided by T.H. Eiermann, University of Ulm, and Red Cross Blood Bank, Ulm, Germany.)

Paternity Testing by RFLP Analysis

Three paternity cases tested with DNA purified from blood samples with the QIAamp DNA Blood Mini Kit. 1: mother; 2: child; 3: alleged father; 4: child + alleged father. M: markers. 3 µg of DNA was loaded per lane. Outcome: A: alleged father excluded; B & C: alleged father confirmed. (Data kindly provided by J. James, Gene Proof Technologies, Nashville, TN, USA.)

* For automated, high-throughput purification from swabs, the QIAamp 96 DNA Swab BioRobot Kit is recommended.
†  The QIAamp 96 Virus BioRobot Kit isolates both DNA and RNA in parallel.