Investigator Quantiplex Kit

 Investigator Quantiplex Kit

Ref.387016

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.

 

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”).

 

Investigator Quantiplex Kit_Spec

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