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Cat. No. / ID: 387216
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Investigator Quantiplex Kits are ready-to-use quantitative real-time PCR assays for human identity testing and forensic applications with conventional 2 µL or flexible 1-18 µL sample input (Investigator Quantiplex Pro FLX Kit). Developed for superior sensitivity and reaction speed, they are designed to establish whether a sample contains sufficient and intact human DNA to enable STR or SNP analysis by Capillary Electrophoresis or NGS. An internal amplification control (IC) is used to test successful amplification and identify the presence of PCR inhibitors. The Investigator Quantiplex Pro technology further enables quality assessment to provide reliable information on the degradation status of your sample.
Investigator Quantiplex Kits meet ISO 18385 requirements.
Investigator Quantiplex Pro Kits provide robust and precise quantification results with high sensitivity within a high dynamic range. In addition to the general sensitivity, the kits provide exceptionally high sensitivity for male DNA, even in a very high background of female DNA (up to 400,000:1).
The straightforward quality assessment system included in the kits comprises a newly developed DNA Degradation Control (DC) and a newly designed Internal Control (IC). The new DNA Degradation Control provides superior detection of DNA degradation (see figures “ Accurate detection of DNA degradation” and “ Accurate detection of male DNA degradation in a sample mixture”). Furthermore, the Investigator Quantiplex Pro RGQ assay is the only commercially available assay that can distinguish between human and male DNA degradation (see figure “ Highly accurate and sensitive detection results for male and total human DNA”). The newly designed Internal Control reliably detects potential PCR inhibitors. The IC is more sensitive to inhibitors and correlates to STR tolerance while the quantification results remain reliable (see figure “ Identification of PCR inhibition”). Furthermore the IC of the Investigator Quantiplex Pro Kits is analogous to the Quality Sensor of Investigator STR QS PCR Assays (e.g., Investigator 24plex QS Kits, Investigator 26plex QS Kits, Investigator ESSplex SE QS Kit and Investigator Argus X-12 QS Kit).
Human DNA isolated from various sources has to be assessed in terms of quantity, quality and integrity prior to STR analyses. STR multiplexes are complex systems that require a narrowly defined range of input DNA and template quality to perform accurately. As DNA quantification is the only step preceding the STR PCR, it is essential to extract as much information as possible from this reaction to aid correct setup of STR reactions.
Investigator Quantiplex Pro Kits are a ready-to-use system for the detection of human and male DNA and parallel assessment of DNA degradation using quantitative real-time PCR. The kits provide fast and accurate quantification of human DNA from forensic database and casework samples.
Detection of amplification is performed using TaqMan probes and a fast PCR chemistry. The dual-labeled probes contain a fluorescent reporter and a quencher at their 5' and 3' ends, respectively. During the extension phase of PCR, the 5' and 3' exonuclease activity of the DNA polymerase cleaves the fluorophore from the quencher. This results in detectable fluorescence that is proportional to the amount of accumulated PCR product. Information on the targets, amplicon lengths, channels, ploidy and copy numbers is summarized in the table “Targets, amplicon lengths and channels for Investigator Quantiplex Kits”.
Kit | Target | Amplicon length | Channel | Ploidy | Copy number |
Quantiplex | Human target | 146 bp | FAM | Diploid | Multi-copy |
Internal PCR control | 200 bp | VIC | N.A. | Synthetic fragment | |
Quantiplex Pro Quantiplex Pro FLX |
Human target, small autos. | 91 bp | FAM | Diploid | Multi-copy |
Human target, large autos. | 353 bp | TAMRA/Atto550 | Diploid | Multi-copy | |
Human male target | 81 bp | CY5/Atto647N | Haploid | Multi-copy | |
Internal PCR control | 434 bp | JOE | N.A. | Synthetic fragment | |
Quantiplex Pro RGQ | Human target, small autos. | 91 bp | Yellow | Diploid | Multi-copy |
Human target, large autos. | 353 bp | Red | Diploid | Multi-copy | |
Human male, small Y | 81 bp | Green | Haploid | Multi-copy | |
Human male, large Y | 359 bp | Orange | Haploid | Multi-copy | |
Internal PCR control | 434 bp | Crimson | N.A. | Synthetic fragment |
The kits are 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. In order to streamline the instrument setup and the analysis of the results, QIAGEN has developed a set of template files. Download the template files from the product resources page.
When using the Applied Biosystems 7500 or QuantStudio 5 Real-Time PCR Systems with Investigator Quantiplex Pro Kit, custom dye calibration for FAM, JOE, ATTO 550, ROX, and ATTO 647N is needed. To achieve optimal performance, calibrate with the Investigator Quantiplex Pro Calibration Kit.
Investigator Quantiplex Kits are for quantifying human and male DNA, and for determining if PCR inhibitors or DNA degradation are present and if further sample purification is needed. They are suitable for:
Technical specifications
All targets are multi-copy targets.
Features | Investigator Quantiplex Pro Kit | Investigator Quantiplex Pro FLX | Investigator Quantiplex Pro RGQ Kit | Investigator Quantiplex Kit |
Volume per PCR | 20 µl | 20 µL | 20 µl | 25 µl |
Input volume of template DNA | 2 µl | 1-18 µL | 2 µl | 2 µl |
Human target, small autosomal | 91 bp | 91 bp | 91 bp | 146 bp |
Human target, large autosomal | 353 bp | 353 bp | 353 bp | - |
Human male target, small Y | 81 bp | 81 bp | 81 bp | - |
Human male target, large Y | - | - | 359 bp | - |
Probes | TaqMan | TaqMan | TaqMan | Scorpions |
Real-time cyclers | Applied Biosystems 7500 Real-Time Systems or QuantStudio 5 Real-Time Systems |
Applied Biosystems 7500 Real-Time System or QuantStudio5 Systems |
Rotor-Gene Q, 5plex or 6plex only |
Rotor-Gene Q, 5plex or 6plex, Applied Biosystems 7500 Real-Time Systems |
Male genomic DNA was sheared with a Covaris S220 Focused-ultrasonicator to average fragment sizes of 1500 bp, 500 bp, 400 bp, 300 bp, 200 bp and 150 bp. [A] Each fragment size (4.6 ng) was tested using the Investigator Quantiplex Pro RGQ Kit. The performance of two other commercially available kits under the same test conditions is shown in [B] and [C]. All reactions were set up and run according to the manufacturer’s instructions. There was a higher shift in CT value for the QIAGEN human L fragment compared to kits from alternative suppliers, indicating a more accurate assessment of DNA quality in the sample. (Data generated by QIAGEN R&D.)