Investigator Quantiplex Pro FLX Kit

For reliable quantification and qualitative assessment of human and male DNA with flexible sample input up to 18 µL

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Want to try this solution for the first time?
Get in touch with our team today and request a quote for your Investigator Quantiplex Pro FLX Kit (576) trial kit.
Investigator Quantiplex Pro FLX Kit (576)

Cat. No. / ID:  387516

For use on Applied Biosystems Real-Time Systems: 6x single blistered 96-well optical PCR plates with Master Mix, Control DNA M1, and QuantiTect Nucleic Acid Dilution Buffer
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KitAccessories
Quantiplex Pro FLX
Quantiplex Pro Calibration
qPCR adhesive plate foil
PCR water
The Investigator Quantiplex Pro FLX Kit is intended for molecular biology applications in forensic, human identity, and paternity testing. This product is not intended for diagnosis, prevention, or treatment of a disease.
Want to try this solution for the first time?
Get in touch with our team today and request a quote for your Investigator Quantiplex Pro FLX Kit (576) trial kit.

Features

  • Ready-to-use 96-well optical plates pre-filled with lyophilized qPCR Mastermix
  • Trusted multicopy markers for advanced assessment of quantity, degradation and inhibition
  • Full flexibility with option to scale input volume from 1 µL to 18 µL for highest sensitivity
  • Breakable plates and convenient seal removal to scale for the throughput needed
  • Sustainable room temperature storage to free-up freezer capacity

Product Details

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.

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

See figures

Performance

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).

 

See figures

Principle

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. 

 

Procedure

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.

 

Applications

The Investigator Quantiplex Pro FLX kit is ideally suited for:

  • Sexual Assault Screening
  • Special samples that requires highest sensitivity
  • High-throughput applications

Room temperature storage makes the kit suitable for mobile labs in disaster victim identification, too.

 

Targets, amplicon lengths and channels for Investigator Quantiplex Pro FLX Kit
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



Technical Specifications
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. 

Supporting data and figures

Resources

Brochures & Guides (4)
Explore our new, easy-to-navigate digital version of the Investigator Quantiplex Pro Kit Product Profile!
Our improved noninvasive paternity testing workflow delivers answers with greater confidence
How do you choose the best downstream analysis path for a critical forensic sample?
Technical Information (2)

Unprecedented flexibility and convenience in forensic quantification


Certificates of Analysis (1)

FAQ

Which targets are used in Investigator Quantiplex Pro FLX?
The targets used are identical to the targets from Investigator Quantiplex Pro Kit (cat. no. 387216). See also the table “Targets, amplicon lengths and channels for Investigator Quantiplex Pro FLX Kit” from the Technical Information Flyer.
FAQ ID - 4027
Can I use the PCR protocol from Quantiplex Pro for Quantiplex Pro FLX?
No, the PCR programs are different. Quantiplex Pro FLX needs higher temperatures for initial Taq activation and denaturation. However, the cycling times are identical for both kits.
FAQ ID - 4035
What are the storage conditions of the Investigator Quantiplex Pro FLX Kit?
The storage temperature for the plates in Box 1 is at room temperature (18–25°C). For the Control DNA M1 and NADB in Box 2, it is 2–8°C. Partially used plates should be stored in the aluminum bag and desiccant at room temperature, and should be used within four weeks.
FAQ ID - 4026
Can I use less than 18 µL sample volume?
Yes, but always fill up with water (e.g., PCR Water (cat. no. 17000-10)) to 18 µL to dissolve the cake. Using less volume is not possible.
FAQ ID - 4033
Why is the color of the lyophilized cake light blue?
The Investigator Quantiplex Pro FLX Kit uses a hot-start DNA polymerase enzyme and QuantiNova Guard additive which further improve the stringency of the antibody-mediated hot-start and features a built-in control for visual identification of correct pipetting.
FAQ ID - 4031
How does the Investigator Quantiplex Pro FLX kit format look like?
The kit comes in two boxes. Box 1 contains six plates with the lyophilized Master mix (single blistered in aluminum bag with desiccant). Box 2 contains the Control DNA M1 as well as the Nucleic Acid Dilution Buffer (NADB).
FAQ ID - 4025
Should I be concerned about the inconsistent cake shape?
The cake shape has no impact on the functionality or performance of the kit.
FAQ ID - 4032
What should I do if the lyophilized cake was accidentally exposed to high humidity?
Add the sample and incubate for 10 minutes at room temperature before trying to completely redissolve the cake.
FAQ ID - 4030
Can I use just a portion of an entire 96-well plate to avoid reagent waste?
Yes, the Investigator Quantiplex Pro FLX chemistry comes in breakable 96-well optical plates. Always break the plates before removing the seal.
FAQ ID - 4028
How should I close the plate after reaction setup?
Always use the qPCR adhesive plate foil, perforated (100) (cat. no. 386055) to close single strips. Use qPCR adhesive plate foil (100) (cat. no. 209003) or an optical adhesive film from a different supplier (e.g., MicroAmp™ Optical Adhesive Film, Thermo Fisher) to close an entire plate. Always use a compression pad (e.g., Optical film compression pad, VWR cat. no. 76637-856) when running full 96-well plates.
FAQ ID - 4034
How should I remove the plate seal to get access to 96 wells at once?
Add 1/3 of an ordinary adhesive film to the side of the removal tapes and peel the seal at once.
FAQ ID - 4029
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 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
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: 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 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 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
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
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
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
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
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
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 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
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