InnoQuant® HY

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InnoQuant® HY – highly sensitive and robust assessment of human and human male DNA quantity, quality and integrity.

  • Highly Informative: includes a Degradation Index (ratio of short to long target quantity), IPC (Internal Positive Control) and Y-chromosome specific male target
  • Highly Sensitive: limit of detection <1 picogram
  • Highly Reproducible: large copy number of selected targets minimizes the effect of variation between individuals
  • Easy to Use: compatible with commonly used real-time PCR instruments & software

Key Benefits:

  • Improves first pass sample success rate
  • Enables confident screening of negative samples
  • Eliminates unnecessary sample retesting
  • Enables selection of optimal downstream typing methods. i.e. autosomal STR, Y-STR or InnoTyper

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InnoQuant HY is a highly sensitive real-time qPCR assay for robust simultaneous assessment of human and male DNA quantity, quality and integrity.

A Better System for Forensic Samples

Forensic samples are highly variable in DNA quantity and quality. Therefore, quantification is a critical step to ensure an appropriate amount of DNA, and downstream typing method, is used for downstream PCR amplification. Many current quantification systems do not fully account for quality differences in forensic samples and can underestimate the optimal amount of amplifiable DNA.

InnoQuant systems provide more comprehensive quality assessment by using two high copy number retrotransposable element targets: a short target (80 bp) for assessment of total DNA quantity, and a long target (207 bp) for better assessment of total amplifiable DNA. The ratio of short to long targets can be used to calculate the Degradation Index to assess the overall quality of a sample. The InnoQuant HY kit also includes a sensitive multi-copy Y-chromosome specific male target for additional information about a sample’s male DNA composition.

Degradation Index vs Peak RFU
Figure 1. The InnoQuant degradation index (DI = short quantity value / long quantity value) accurately assesses degradation in a forensic sample. As the DI increases, the peak height (RFU) of STR alleles in downstream profiling decreases.

Highly Sensitive and Reproducible

InnoQuant systems are highly sensitive, reproducible and accurate due to the high copy nature of the assay targets. As shown in the figure below, the InnoQuant HY kit can consistently detect sub-picogram amounts of DNA. This enables you to confidently screen samples and avoid needless processing of negative samples.

Degradation Index vs Peak RFU
Figure 2. A serial dilution of the NIST SRM 2372-A Human DNA Quantitation Standard was quantified in triplicate from neat solutions diluted to 0.078125 pg/µL, and mean concentrations compared to values listed in the SRM certificate.

In addition, the concentration of the pooled genomic male DNA used for the InnoQuant HY DNA Standard is calibrated against the NIST SRM 2372-A Human DNA Quantitation Standard to increase lot-to-lot consistency.

Adjust DNA Target to Improve Success Rates with Challenging Samples

When utilizing both the total DNA quantity and the Degradation Index, forensic analysts can better assess the optimal amount of DNA to amplify. Adjusting the amount of input DNA, on even moderately degraded samples, can greatly improve DNA profiles. As shown in the figure below, increasing the DNA input based on the InnoQuant long target results led to 130% more STR alleles being called.

Using InnoQuant Can Improve Profile Success
Figure 3. Increasing the DNA input based on the Degradation Index and the InnoQuant long target results led to 130% more alleles being called. (Allele detection measured using Identifiler® Plus PCR Amplification Kit and Applied Biosystems 3130 Genetic Analyzer.) Use of both the Degradation Index and the quantity values can improve STR allele recovery and reliably predict downstream STR success.

Accurate Assessment of Male-Female Mixtures

With the inclusion of a sensitive multi-copy Y-chromosome specific male target, InnoQuant HY enables you to accurately determine male DNA quantity in challenging male-female mixtures (i.e. sexual assault samples), and more easily identify when sample processing should be stopped, modified, or if the sample should be preserved for alternative testing methods such as InnoTyper or Y-STR testing. As shown in the figure below, male DNA can be accurately detected in samples with large female DNA concentrations.

Figure 4 Mixture Study
Figure 4. Mean quantification results from a series of samples run in triplicate that were prepared with mixtures of human male (NIST SRM 2372-A) and human female (NIST SRM 2372-B) DNA. The male DNA was kept at a constant level of 10 pg/µL while the female DNA was added at increasing levels to create the male-to-female ratio range from 1:1 to 1:8000. The InnoQuant HY kit was able to detect male DNA up to the highest ratio of 1:8000 tested. Additional studies conducted by a third party laboratory have demonstrated the kit’s ability to detect as little as 1 pg/µL of male DNA in a male-to-female ratio of approximately 1:50,000 (data not shown).

Improve Workflow and Eliminate Sample Retesting

A quantification method that provides more comprehensive information on sample quality can significantly improve the overall workflow in your laboratory. Better decisions can be made on which samples to amplify, the appropriate DNA target to amplify, and whether to concentrate or dilute a sample prior to amplification. This added information allows for a more streamlined workflow and minimizes unnecessary sample processing and retesting.

InnoQuant HY enables you to:

  • Confidently screen negative samples
  • Improve first pass success rate
  • Eliminate unnecessary sample retesting
  • Select optimal protocols and typing test kits
  • Identify samples to preserve for alternative methods such as InnoTyper or Y-STR testing

COMPATIBLE WITH COMMONLY USED REAL-TIME PCR INSTRUMENTS & SOFTWARE

With multiple dye configuration options, the InnoQuant HY kit is fully compatible with the Applied Biosystems® 7500 Real-Time PCR System, the BioRad CFX Real-Time PCR System, and other instruments. Costly instrument or software upgrades are not required.

Figure 4 Mixture Study
Figure 5. Dye configurations for the InnoQuant HY and InnoQuant HY-R kits. The InnoQuant HY kit is recommended for use with the Applied Biosystems® 7500 Real-Time PCR System, while the InnoQuant HY-R kit is recommended for other RT-PCR instrument platforms that do not require the use of a reference dye.
Figure 4 Mixture Study
Figure 6. Amplification plots of the DNA Standards and NTC for short (A), long (B), Y (C) and IPC (D) targets.
Figure 4 Mixture Study
Figure 7. Standard Curves of the short, long and Y targets using DNA standard dilutions ranging from 20 ng/µL to 0.005 ng/µL.

For research use only. Not for use in diagnostic procedures.

Protocols & Technical Resources

Software Templates

Publications

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Symposium Presentations & Posters

Product Size Components Catalog #
InnoQuant HY Human and Male DNA Quantification & Degradation Assessment Kit 250 reactions DNA Standard – 75 µl @ 100 ng/µl
Primer Mix – 2 tubes, 1.00 mL each
Dilution Buffer A – 2 tubes, 1.80 mL each
Agilent Technologies Brilliant Multiplex QPCR Master Mix – 2.5 mL
ROX Reference Dye - 100 μL at 1 mM
21108-250 Request a Quote
InnoQuant HY Human and Male DNA Quantification & Degradation Assessment Kit 60 reactions DNA Standard – 33 µl @ 100 ng/µl
Primer Mix – 1 tube, 0.5 mL each
Dilution Buffer A – 1 tube, 1.80 mL each
Agilent Technologies Brilliant Multiplex QPCR Master Mix – 0.5 mL
ROX Reference Dye - 100 μL at 1 mM
21108-060 Request a Quote
InnoQuant HY-R Human and Male DNA Quantification & Degradation Assessment Kit (for non-7500 instruments) 250 reactions DNA Standard – 75 µl @ 100 ng/µl
Primer Mix – 2 tubes, 1.00 mL each
Dilution Buffer A – 2 tubes, 1.80 mL each
Agilent Technologies Brilliant Multiplex QPCR Master Mix – 2.5 mL
ROX Reference Dye - 100 μL at 1 mM
21104-250 Request a Quote
InnoQuant HY-R Human and Male DNA Quantification & Degradation Assessment Kit (for non-7500 instruments) 60 reactions DNA Standard – 33 µl @ 100 ng/µl
Primer Mix – 1 tube, 0.5 mL each
Dilution Buffer A – 1 tube, 1.80 mL each
Agilent Technologies Brilliant Multiplex QPCR Master Mix – 0.5 mL
ROX Reference Dye - 100 μL at 1 mM
21104-060 Request a Quote
InnoQuant Spectral Calibration Kit 33 uL of each dye at 100 uM Calibration standards for HEX, Cy3, Cy5, and TAMRA 21105 Request a Quote

 

Storage Conditions: Upon receipt, store the kit at -20°C. After thawing, store all components at 2°C to 8°C.