For rapid and reliable quantification of human DNA in forensic samples

  • PCR speed of 48 minutes on the Rotor-Gene Q
  • Reliable results with sensitivity down to <1 pg/µl DNA
  • Integrated internal control to test for PCR amplification or inhibition
  • Fully automatable reaction setup

The Investigator Quantiplex Kit is a quantitative real-time PCR assay for human identity and forensic testing applications. Developed for superior sensitivity and reaction speed, it is designed to establish whether a sample contains sufficient DNA to enable DNA fingerprinting analysis (such as STR or DIP analysis) and whether there are inhibitors present.

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Investigator Quantiplex Kit (200)
Primer Mix IC FQ, Reaction Mix FQ, Control DNA Z1, Dilution Buffer
387016
$481.00
The Investigator Quantiplex Kit is intended for molecular biology applications in forensic, human identity, and paternity testing. This product is not intended for diagnosis, prevention, or treatment of a disease.
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The Investigator Quantiplex Kit retains excellent sensitivity down to 1 pg/µl.|Superior reaction time.|No copy-number variation in the marker.|Scorpions primers and their function.|
Control DNA Z1 was diluted to 0.3 pg/μl and quantified using the Investigator Quantiplex Kit in 2 independent runs.|The Investigator Quantiplex Kit is significantly faster than similar kits from other suppliers on both the Rotor-Gene Q and Applied Biosystems 7500 Fast Real-Time PCR System.|A quantitative real-time PCR was run to compare a specific single-copy human DNA target and the Investigator Quantiplex multi-copy target. The ΔCT between the two targets is shown for the different populations.||
Performance

The validation of Investigator Quantiplex Kit showed that it yielded robust and reproducible results within the normal range of conditions expected in forensic casework and human identity testing. The kit enables a significantly faster PCR than similar kits from other suppliers, yielding results in just 48 minutes on QIAGEN's Rotor-Gene Q thermal cycler (see the figure "Superior reaction time").

The marker of the Investigator Quantiplex Kit has been validated and no copy-number variation has been detected between genders or across populations, making this marker an ideal real-time PCR target for DNA quantification (see the figure "No copy-number variation in the marker").

Furthermore, the assay provides sensitivity down to <1 pg/µl, with accurate quantification of 4.9 pg/µl, where the standard curve shows linearity (see the figure "The Investigator Quantiplex kit retains excellent sensitivity down to 1 pg/µl").

Principle

Human identification is commonly based on the analysis of short tandem repeats (STRs), single nucleotide polymorphisms (SNPs), or deletion insertion polymorphisms (DIPs), depending on the demands of an examination or on the sample quality. These multiplex assays are complex systems that require a defined range of template input.

The Investigator Quantiplex Kit contains reagents and a DNA polymerase for specific amplification of a 146 bp autosomal multi-copy region of the human genome and for direct detection of the specific PCR product on the Rotor-Gene Q or Applied Biosystems 7500 Real-Time PCR Systems. The proprietary multi-copy region was selected in order to give high sensitivity with high reliability within different genders and populations. The target is detected using the green channel on the Rotor-Gene Q or the FAM dye channel on Applied Biosystems instruments.

In addition, the Investigator Quantiplex Kit contains a balanced internal amplification control that is used to test successful amplification and detects PCR inhibitors. This heterologous amplification system is detected as a 200 bp internal control (IC) in the yellow channel on the Rotor-Gene Q or in the VIC dye channel on Applied Biosystems instruments.

Detection of amplification is performed using Scorpions primers and a novel, fast PCR chemistry. Scorpion primers are bifunctional molecules containing a PCR primer covalently linked to a probe (see the figure "Scorpions primers and their function"). The fluorophore in this probe interacts with a quencher, also incorporated into the probe, which reduces fluorescence. During PCR, when the probe binds to the amplicon, the fluorophore and quencher become separated. This leads to an increase in fluorescence in the reaction tube.

The reaction chemistry was carefully optimized to further support the rapid mechanism. The novel PCR chemistry and enzyme, which were specifically developed for human identification, enable high sensitivity and speed. Furthermore, the specially developed PCR chemistry contains the patent-pending additive Q-Bond, which significantly reduces denaturation, annealing, and extension times.

Procedure

The kit is a ready-to-use system for the detection and quantification of human DNA using real-time PCR. See the table "Technical specifications" for more information. A set of template files has been developed to streamline the instrument setup and the analysis of the results on the Rotor-Gene Q (see table "Template files available for the Rotor-Gene Q").

Technical specifications
Volume per PCR reaction 25 µl
Input volume of template DNA 2 µl
Target  Human-specific genomic DNA multi-copy target
Real-time cyclers Rotor-Gene Q, Applied Biosystems 7500 Real-Time System, or
Applied Biosystems 7500 Fast Real-Time System
Template files available for the Rotor-Gene Q
Template filePurpose
Investigator Quantiplex Kit Cycling Preset cycling protocol
Investigator Quantiplex Kit Sample Sample name, sample type and concentration for standard curve and internal control
Investigator Quantiplex Kit Analysis Settings for the Yellow Channel CT settings
Applications

The Investigator Quantiplex Kit is for quantifying human DNA, and for determining if PCR inhibitors are present and if further sample purification is needed. It is suitable for:

Forensic database and casework samples
Paternity testing samples
Disaster victim identification and anthropological work
Other human identity testing applications

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