Description
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”. 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 N.A. = Not applicable
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.
