Investigator Quantiplex Kit

For reliable quantification and qualitative assessment of human and male DNA with conventional 2 µL sample input

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Investigator Quantiplex Pro Kit (200)

Cat. No. / ID:  387216

For use on Applied Biosystems Real-Time Systems: Quantiplex Pro Reaction Mix, Quantiplex Pro Primer Mix, Control DNA M1, QuantiTect Nucleic Acid Dilution Buffer
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Quantiplex Pro
Quantiplex Pro RGQ
Quantiplex
Quantiplex Pro Calibration
qPCR adhesive plate foil
Investigator Quantiplex Kits are intended for molecular biology applications in forensic, human identity and paternity testing. These products are not intended for the diagnosis, prevention or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Simultaneous quantification of total human or human and male DNA
  • Straightforward quality assessment for reliable information on DNA degradation correlating with your STR profiles
  • Reliable results with sensitivity down to <1 pg/µL DNA
  • High sensitivity for male DNA in a female background
  • Integrated internal control to test for PCR amplification or inhibition

Product Details

Investigator Quantiplex Kits are ready-to-use quantitative real-time PCR assays for human identity testing and forensic applications with conventional 2 µL sample input. 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.

Want to try the Investigator Quantiplex Pro Kit for the first time? Request a quote for a trial kit.

Want to try the Investigator Quantiplex Pro RGQ Kit for the first time? Request a quote for a trial kit.

Performance

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).

See figures

Principle

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 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

Procedure

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.

Applications

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:

  • Forensic database and casework samples
  • Paternity testing samples
  • Sexual assault samples
  • Disaster victim identification and anthropological work
  • Other human identity testing applications

Technical specifications

All targets are multi-copy targets.

Features Investigator Quantiplex Pro Kit Investigator Quantiplex Pro RGQ Kit Investigator Quantiplex Kit
Volume per PCR 20 µl 20 µl 25 µl
Input volume of template DNA 2 µl 2 µl 2 µl
Human target, small autosomal 91 bp 91 bp 146 bp
Human target, large autosomal 353 bp 353 bp -
Human male target, small Y 81 bp 81 bp -
Human male target, large Y - 359 bp -
Probes TaqMan TaqMan Scorpions
Real-time cyclers Applied Biosystems 7500 Real-Time Systems or QuantStudio 5 Real-Time Systems

Rotor-Gene Q, 5plex or 6plex only

Rotor-Gene Q, 5plex or 6plex, Applied Biosystems 7500 Real-Time Systems

 

Supporting data and figures

Resources

Brochures & Guides (4)
Our improved noninvasive paternity testing workflow delivers answers with greater confidence
Explore our new, easy-to-navigate digital version of the Investigator Quantiplex Pro Kit Product Profile!
How do you choose the best downstream analysis path for a critical forensic sample?
Scientific Posters (4)
Galina Kulstein, Thorsten Hadrys, Keith Elliott, Miro Vranes and Peter Wiegand. Scientific Poster presented at ISHI 2017, Seattle, Washington, USA and kindly provided by Galina Kulstein, Institute of Legal Medicine, DNA Department, University Hospital of Ulm, Germany and published with permission.
Kit Handbooks (4)
For quantification of human and male DNA in forensic samples
For quantification of human DNA in forensic samples
For quantification of human and male DNA in forensic samples
For quantification of human and male DNA in forensic samples
Technical Information (4)
Certificates of Analysis (1)

FAQ

Investigator Quantiplex: Which version of the Applied Biosystems 7500 Real-Time PCR System software can be used to run the Investigator Quantiplex?
On the Applied Biosystems 7500 Real-Time PCR System, use SDS software version 1.4.
FAQ ID -2569
Investigator Quantiplex: Do I have to calibrate new dyes on my Applied Biosystems 7500 or 7500 Fast Real-Time PCR System?
No, you do not. Please use the FAM and the VIC dyes as explained in the Investigator QuantiPlex Kit handbook.
FAQ ID -2571
Which human target is used for the quantification in the Investigator Quantiplex Kit?
For all Investigator Quantiplex Kits, the 4NS1C target is used, a multi-copy target which was validated in an external study. It is represented by about 20 copies per haploid genome and is located on different autosomal chromosomes.
FAQ ID -2577
Investigator Quantiplex Pro: On which real-time cyclers is the Investigator Quantiplex Pro Kit validated?
The Investigator Quantiplex Pro Kit is validated on the Applied Biosystems 7500 Standard, Applied Biosystems 7500 Real-Time PCR System for Human Identification, and QuantStudio 5 Real-Time HID PCR Systems.

FAQ ID - 3728
Investigator Quantiplex and Quantiplex Pro: What do I have to consider if I am using Applied Biosystems SDS software version 1.2.3?
If using software version 1.2.3, be sure to uncheck the box “9600 Emulation” in the cycling set-up.
FAQ ID -2570
How long can I store the dilutions of the Male Control DNA M1 standards at 2–8°C?
If the series of the Male Control DNA M1 standards was diluted using the QuantiTect Nucleic Acid Dilution Buffer provided, then they can be stored for at least one week at 2–8°C. 
FAQ ID - 3744
Why do I observe a CT shift for the Internal Control in some samples in comparison to the CT value for the Internal Control of the dilution series of the Male Control DNA M1?
The Internal Control is designed to report if inhibitors are contained in the DNA sample. A shift in the CT value in comparison to the dilution series of the Male Control DNA M1 is an indication of potential inhibition in the DNA sample.
FAQ ID - 3745
Investigator Quantiplex Pro: Which real-time cycler software version can be used to run the kit?

Using the Investigator Quantiplex Pro Kit on the Applied Biosystems 7500 Real-Time PCR System:
All software versions starting with SDS v. 1.2.3 can be used. If you are using software version 1.2.3, ensure that you uncheck the box 9600 Emulation in the cycling setup.

FAQ ID - 3730
Investigator Quantiplex Pro RGQ: Which real-time cycler software version can be used to run the kit?

Using the Investigator Quantiplex Pro RGQ Kit on the Rotor-Gene Q System:

all software versions starting from Q-Rex 1.0 can be used.
FAQ ID - 3743
Investigator Quantiplex Pro RGQ: Which Rotor-Gene Q System can be used to run the kit?
The Investigator Quantiplex Pro RGQ Kit can only be run on the Rotor-Gene Q System (5plex/6plex) with 72-Well Rotor/Rotor-Disc 72/Rotor-Disc 72 Rotor and Rotor-Disc 100/Rotor-Disc 100 Rotor.
FAQ ID - 3742
Why is the fluorescence signal of the Internal Control on my Rotor-Gene Q very low?
Be sure to perform fluorescence data collection using the green and the yellow channels with auto-gain optimization.
FAQ ID -2574
Investigator Quantiplex Pro: Do I have to calibrate new dyes on my Applied Biosystems 7500 or QuantStudio 5 Real-Time PCR System?
With the Investigator Quantiplex Pro Kit, custom dye calibration for FAM, JOE, ATTO 550, ROX, and ATTO 647N is strongly recommended when using the Applied Biosystems 7500 or Quantstudio 5 Real-Time PCR Systems. To achieve optimal performance, please calibrate with the Investigator Quantiplex Pro Calibration Kit (cat. no. 387416). However, standard dye calibrations for FAM, JOE, TAMRA, ROX, and Cy5 might be used. In this case, use TAMRA for the Degradation target and Cy5 for the Male target.
FAQ ID - 3732
How long does a run take?
An Investigator Quantiplex Pro or Quantiplex Pro RGQ run of 40 cycles takes ca. 60 minutes. 
For Investigator Quantiplex on the Rotor-Gene Q, a run of 40 cycles takes ca. 55 minutes. On the Applied Biosystems 7500 Real-Time PCR System, a run of 40 cycles takes ca. 57 minutes.
FAQ ID -2566
Investigator Quantiplex: For which real-time cyclers has the kit been validated?
The Investigator Quantiplex Kit has been validated for the Rotor-Gene Q, and the Applied Biosystems 7500 Fast and 7500 Real-Time PCR Systems.
FAQ ID -2567
What are the storage conditions?
The Investigator Quantiplex Pro and Investigator Quantiplex Pro RGQ Kit is stored at –15°C to –30°C. After first time use, store kit components at 2–8°C. Avoid freezing the kit components.  The Investigator Quantiplex Kit is stored at 2–8°C. The QuantiTect Nucleic Acid Dilution Buffer may also be stored at –15°C to –30°C, if desired. 
FAQ ID - 3748
Investigator Quantiplex: Which version of the Rotor Gene-Q software can be used to run the kit?
Using the Investigator Quantiplex on the Rotor-Gene Q: All versions of the Rotor-Gene Q software can be used to run the Investigator Quantiplex.
FAQ ID -2568
How can I streamline the quantification workflow?
You can download the template files for the corresponding assay from the website and use them according to the handbook. These files will allow you to use the right parameters to run a quantification reaction and analyze the results in the proper way.
FAQ ID - 3738
Investigator Quantiplex Pro: Can the kit be run on the Rotor-Gene Q?
No, the dyes are not compatible with the Rotor-Gene Q. A separate Rotor-Gene Q version is available:
Investigator Quantiplex Pro RGQ Kit (200) (Cat. No./ID: 387316)
FAQ ID - 3729
Which human target is used for the quantification?
The 4NS1C target is a multi-copy target, which was validated in an external study. It is located on different autosomal chromosomes.
FAQ ID - 3747
Investigator Quantiplex Pro RGQ: On which real-time cyclers is the kit validated?
The Investigator Quantiplex Pro RGQ Kit is validated on the Rotor-Gene Q System.
FAQ ID - 3741