Investigator Quantiplex HYres Kit

For detection and quantification of human and male DNA in forensic samples

  • Simultaneous quantification of total human and male DNA
  • Reliable results with sensitivity down to <1 pg/µl DNA
  • PCR speed of 51 minutes on the Rotor-Gene Q
  • Intergrated internal control to test for PCR amplification or inhibition
  • Fully automatable reaction setup

The Investigator Quantiplex HYres Kit is a ready-to-use system for the detection and quantification of total human and male DNA using real-time PCR. 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. The Investigator Quantiplex HYres Kit provides reliable results independent of the subject's gender or ethnic background.

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Investigator Quantiplex HYres Kit (200)
Reaction Mix FQ, Primer Mix IC YQ, Control DNA Z1, QuantiTect Nucleic Acid Dilution Buffer
387116
752,00 €
The Investigator Quantiplex HYres 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.|Excellent reaction time.|No copy-number variation in the human target.|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 HYres Kit yields results in 51 minutes on the Rotor-Gene Q and 62 minutes on the Applied Biosystems 7500 Real-Time PCR Cycler. This is significantly faster than kits from other suppliers.|The human genetic target of the Investigator Quantiplex HYres Kit is highly conserved and does not vary across ethnicity. The change in threshold cycle between the two targets is shown across populations. n equals the number of individuals in the test group.||
Performance

The validation of Investigator Quantiplex HYres 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 51 minutes on QIAGEN's Rotor-Gene Q thermal cycler (see the figure "Excellent reaction time"). This enables a time- and cost-effective workflow.

The markers 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 human target").

The kit can quantify a broad range of DNA amounts, from 50 ng/µl to about 0.5 pg/µl with a highly linear range between 20 ng/µl and 4.9 pg/μl of human genomic DNA. Furthermore, the assay provides sensitivity down to <1 pg/μl(see the figure "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 HYres Kit contains reagents and a DNA polymerase for the specific amplification of two target regions and the direct detection of the specific PCR products on the Rotor-Gene Q real-time PCR cycler or Applied Biosystems 7500 Real-Time PCR Systems. The target for total human DNA is a 146 bp autosomal multi-copy region of the human genome. This quantification target was selected in order to give high sensitivity with high reliability within different individuals and populations. It is detected using the green channel on the Rotor-Gene Q or the FAM dye channel on Applied Biosystems instruments.

The target region for male DNA quantification was selected in order to reliably give the same high sensitivity within different individuals and populations and in the presence of mixed DNA samples. This target region is detected as a 129 bp fragment using the red channel on the Rotor-Gene Q or the Cy5 dye channel on Applied Biosystems instruments.

In addition, the Investigator Quantiplex HYres Kit contains a balanced internal amplification control that is used to test successful amplification and identify the presence of 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. Scorpions 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 and male DNA 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 20 µl
Input volume of template DNA 2 µl
Target  Human-specific genomic DNA multi-copy target
Male-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 HYres Kit Cycling Preset cycling protocol
Investigator Quantiplex HYres Kit Sample Sample name, sample type and concentration for standard curve and internal control
Investigator Quantiplex Kit HYres Analysis Settings for the Yellow Channel CT settings
Applications

The Investigator Quantiplex HYres kit is for quantifying total human and male 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
Sexual assault samples
Disaster victim identification and anthropological work
Other human identity testing applications

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Kit Handbooks
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For quantification of human and male DNA in forensic samples
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