Cat. No. / ID: 387516
The Investigator Quantiplex Pro FLX Kit is the next generation of quantification in human identification testing. The kit combines trusted multicopy markers for advanced assessment of quantity, degradation and inhibition, with novel lyophilized chemistry in 96-well breakable optical plates (see figure Investigator Quantiplex Pro FLX features: Ease-of-use). It enables for more flexibility in template input volumes starting from 1 µL to up to 18 µL per reaction, and thus offers increased sensitivity by higher input volumes. Purified DNA or sample lysates processed with the Investigator Casework GO! Kit can be used for qPCR. The high template input volumes and fast reaction setup times make the new Investigator Quantiplex Pro FLX Kit ideal for high throughput application and screening approaches where highest sensitivity is pivotal. The kit is validated on Applied Biosystems 7500 or QuantStudio 5 Real-Time PCR Systems.
Investigator Quantiplex Pro FLX meet ISO 18385 requirements.
With the increase of sample template, Investigator Quantiplex Pro FLX Kit provides robust and precise quantification results with highest sensitivity within a high dynamic range. In addition to the 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 9-times higher input volumes lead to approximately 10-times higher sensitivity. This is demonstrated by serial dilution of male DNA (see figure Highest sensitivity by higher input volumes), samples isolated from postcoital swabs and mock samples with semen dilutions (see figure Higher sensitivity for sexual assault sample screening ) as well as low level-DNA casework samples from different sample types like blood, saliva and semen (see figure Low level-DNA casework samples).
Human DNA isolated from various sources has to be assessed in terms of quantity, quality and integrity prior to STR analyses. Either for CE or NGS based methods, these 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. The new Investigator Quantiplex Pro FLX quantification kit combines trusted multicopy markers for advanced assessment of quantity, degradation and inhibition, with novel lyophilized chemistry in 96-well breakable optical plates. The lyophilized chemistry enables more flexibility in template input volumes up to 18 µL per reaction.
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 targets, amplicon length and fluorophore used can be found from the table below.
Investigator Quantiplex Pro FLX is a ready-to-use system for the detection and quantification of human DNA using real-time PCR. The chemistry is pre-filled as lyophilized mastermix in a 96-well optical plate which can be broken into 8-well strips. Removal tabs and seal perforation help to peel the foil from “broken” plates. Alternatively, just add 1/3 of a standard adhesive film to easily peel the seal at once. Template and standards can be added manually or automatically. For both setups, seal the plates with an optical adhesive foil, vortex and centrifuge briefly.
When using the Applied Biosystems 7500 or QuantStudio 5 Real-Time PCR Systems with Investigator Quantiplex Pro FLX, 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.
The Investigator Quantiplex Pro FLX kit is ideally suited for:
Room temperature storage makes the kit suitable for mobile labs in disaster victim identification, too.
Target | Amplicon length | Channel | Ploidy | Copy number |
Human target, small autosomal (Human) | 91 bp | FAM | Diploid | Multi-copy |
Human target, large autosomal (Degradation) | 353 bp | ATTO 550/TAMRA | Diploid | Multi-copy |
Human male target (Male) | 81 bp | ATTO 647N/CY5 | Haploid | Multi-copy |
Internal PCR Control | 434 bp | JOE | N.A. | Synthetic fragment |
Chemistry Format | Lyophilized Master Mix in 96-well optical plate |
Probe | TaqMan |
Template Volume | 1-18 µL |
Volume per PCR | 20 µL* |
Lyophilized cake + 18 µL input volume will yield a final PCR reaction volume of 20 µL.
The Investigator Quantiplex Pro FLX Kit offers unprecedented flexibility and convenience. Breakable optical plates and perforated seals help to support the throughput needed, whilst avoiding reagent waste.