Real-time PCR, also known as quantitative PCR or qPCR, is a powerful technique that allows the detection and quantification of DNA in a sample. This methodology is widely used for various human identification applications, such as forensic databases, forensic casework, confirmatory screening, sexual assault sample workflow, quality, and quantity sample assessment, etc. Real-time PCR can provide accurate, sensitive, robust, and fast results that can help solve crimes, identify victims, and establish paternity.
QuantStudio 5: A Reliable and Affordable Real-Time PCR System for Human Identification
However, not all real-time PCR systems are created equal. Some systems may be too complex, too expensive, or too unreliable for the needs of human identification laboratories. That’s why Applied Biosystems developed the QuantStudio 5 Real-Time PCR System for Human Identification (HID), a new product that offers a simple, reliable, and affordable solution for real-time PCR. The QuantStudio 5 HID system is designed for both new and experienced users who need an easy-to-use, reliable, and affordable real-time PCR system that doesn’t require compromise on performance or quality.
The QuantStudio 5 HID system has many features and benefits that make it an ideal choice for human identification applications. In this article, we will explore some of these features and benefits in detail, and show you how the QuantStudio 5 HID system can help you achieve your human identification goals.
QuantStudio 5 Features and Benefits
Streamlined Design
The QuantStudio 5 HID system has a streamlined design that saves bench space, simplifies operation, and enhances user experience. The QuantStudio 5 system has a small footprint of 27 x 50 x 40 cm (W x D x H), which means it can fit easily on any laboratory bench. The system also has a front-facing fully motorized block that allows easy access to the samples and reduces the risk of contamination. The system also has a touch screen interface that enables viewing of amplification plots, easy and intuitive interface, and USB connectivity.
The streamlined design of the QuantStudio 5 HID system makes it easy to set up, run, and monitor your real-time PCR experiments. You can use the touch screen interface to design your experiments, select your parameters, and start your runs. You can also view your amplification plots in real time on the screen, or export them to a USB drive or a computer for further analysis. The touch screen interface also allows you to access various settings and options, such as calibration, maintenance, diagnostics, etc.
Interactive Software
The QuantStudio 5 HID system comes with interactive software that helps you design, run, and analyze your real-time PCR experiments with ease and flexibility. The software includes the QuantStudio 5 Design and Analysis Software (DAS) and the HID Real-Time PCR Analysis Software (HID RAS).
The QuantStudio 5 DAS is a desktop software that allows you to create and edit your experiments on your computer before transferring them to the instrument. The QuantStudio 5 software also allows you to analyze your data after the run is complete. The software supports various analysis methods, such as standard curve, relative quantification, allelic discrimination, high-resolution melting (HRM), etc. The software also allows you to generate various reports and graphs to visualize and interpret your results.
The HID RAS is a cloud-based software that allows you to access your data from anywhere using a web browser. Human identification applications, such as forensic DNA quantification, are the exclusive focus of the software’s design. Quantifiler Trio, HP, and other Applied Biosystems quantification kits are supported by the software. The software lets you check your samples for male/female ratio, inhibition level, degradation level, etc. To visualise and explain data, the software allows you to generates reports and graphs.
The interactive software of the QuantStudio 5 HID system gives you more control over your real-time PCR experiments. You can use the software to customize your experiments according to your needs and preferences. You can also use the software to analyze your data in different ways and generate meaningful insights from your results.
Reliable Performance
The QuantStudio 5 HID system delivers reliable performance that ensures consistent results, long-term durability, data storage, and quality assurance. The system has several features that contribute to its reliability:
- Bright white light-emitting diode (LED) source with a median lifetime of >5 years
- Less frequent maintenance
- 10 GB on-board memory for storage of 2,000–5,000 run files
- HID-validated system
The bright white LED source provides uniform illumination across the entire block and reduces optical variation between wells. The LED source also has a long lifespan that reduces the need for replacement or calibration. The LED source also enables multiplexing capability for up to six dyes per well, which allows you to detect more targets in a single reaction.
The QuantStudio 5 HID system requires less frequent maintenance than other real-time PCR systems. Dust and other particles are cleared from the optical path using the system’s self-cleaning feature. Additionally, the system includes a self-diagnostic tool that warns you of any flaws or prospective problems. The device also includes a calibration mechanism that enables you to calibrate it whenever necessary using various dyes.
The QuantStudio 5 HID system
The system has a large on-board memory that allows you to store your run files on the instrument. This means you don’t have to worry about losing your data or transferring it to another device. You can access your run files from the touch screen interface or from the DAS software. You can also export your run files to a USB drive or a computer for backup or further analysis.
The QuantStudio 5 HID system is a HID-validated system that meets the quality standards and specifications for human identification applications. The system has been tested and verified with various quantification kits from Applied Biosystems, such as Quantifiler Trio DNA Quantification Kit, Quantifiler HP DNA Quantification Kit, etc. The system also complies with various regulatory and accreditation requirements, such as ISO 17025, SWGDAM, etc.
The reliable performance of the QuantStudio 5 HID system gives you confidence in your results and peace of mind in your workflow. You can trust the system to deliver accurate, consistent, and robust results that can help you solve cases, identify individuals, and establish relationships.
Innovative Technology
The QuantStudio 5 HID system incorporates an innovative technology that enables future quantification technology, optimizes reaction conditions, and customizes thermal profiles. The system has several features that demonstrate its innovation:
- Multiplexing capability for up to six dyes per well
- Precise temperature control with Peltier-controlled blocks
- VeriFlex zones
The multiplexing capability of the QuantStudio 5 HID system allows you to detect more targets in a single reaction, which can save time, cost, and sample. The system can detect up to six dyes per well, which covers the full spectrum of fluorescence from blue to red. The system also has six decoupled filters that can be combined in up to 21 combinations, which gives you more flexibility and versatility in your dye selection.
The precise temperature control of the QuantStudio 5 HID system allows you to optimize your reaction conditions and improve your results. The system uses Peltier-controlled blocks that can heat and cool rapidly and uniformly across the entire block. The system also has a fast ramp rate of 6.5°C/sec and an average sample ramp rate of 3.66°C/sec, which means you can complete your runs faster and more efficiently.
The VeriFlex zones of the QuantStudio 5 HID system allow you to customize your thermal profiles and create different temperature zones within the same block. The system has six independent temperature zones that can be set at different temperatures within a range of ±15°C from the block temperature. This means you can run different experiments or optimize different parameters in the same run.
The innovative technology of the QuantStudio 5 HID system gives you more options and possibilities in your real-time PCR experiments. You can use the system to explore new quantification technologies, fine-tune your reaction conditions, and create different thermal profiles.
QuantStudio 5 Applications
The QuantStudio 5 HID system can be used for various human identification applications, such as forensic databases, forensic casework, confirmatory screening, sexual assault sample workflow, quality and quantity sample assessment, etc. Here are some examples of how the system can help you achieve your human identification goals:
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Forensic database:
If you work in a forensic database laboratory without direct amplification workflows, you need to quantify and qualify your DNA samples before amplifying them with STR kits. The QuantStudio 5 HID system can help you perform accurate and sensitive DNA quantification using Applied Biosystems quantification kits, such as Quantifiler Trio DNA Quantification Kit or Quantifiler HP DNA Quantification Kit.
These kits can measure total human DNA, male DNA, internal positive control (IPC), degradation index (DI), and inhibition index (II) in a single reaction. The kits also have multiplexing capability for up to six dyes per well, which means you can use less sample and reagents per reaction. The QuantStudio 5 HID system can also help you perform quality checks on your samples using the HID RAS software, which can calculate male/female ratio, inhibition level, degradation level, etc. These quality checks can help you determine the optimal amount of DNA input and PCR cycles for STR amplification.
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Forensic casework:
If you work in a forensic casework laboratory, you need to analyze various types of samples from crime scenes or victims, such as blood, saliva, semen, hair, etc. The QuantStudio 5 HID system can help you perform confirmatory screening of male DNA using Applied Biosystems Yfiler Plus PCR Amplification Kit, a 6-dye multiplex assay that amplifies 27 Y-STR loci in a single reaction. The Yfiler Plus kit includes seven rapidly-mutating (RM) Y-STRs that can help resolve paternal lineage differentiation and discriminate closely-related males.
The Yfiler Plus kit also has a high sensitivity and robustness that can help recover alleles from degraded or inhibited samples. The QuantStudio 5 HID system can also help you perform sexual assault sample workflow using Applied Biosystems Quantifiler Duo DNA Quantification Kit, a 5-dye multiplex assay that measures total human DNA, male DNA, and internal positive control (IPC) in a single reaction. The Quantifier Duo kit can help you determine the presence and amount of male DNA in a female background, which can help you select the appropriate STR kit for downstream analysis.
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Quality and quantity sample assessment:
If you need to assess the quality and quantity of your DNA samples before performing STR analysis, the QuantStudio 5 HID system can help you perform accurate and reliable DNA quantification using Applied Biosystems quantification kits, such as Quantifier Trio DNA Quantification Kit or Quantifiler HP DNA Quantification Kit. These kits can measure total human DNA, male DNA, internal positive control (IPC), degradation index (DI), and inhibition index (II) in a single reaction. These parameters can help you evaluate the quality and quantity of your samples and optimize your STR analysis conditions. The QuantStudio 5 HID system can also help you perform quality checks on your samples using the HID RAS software, which can calculate the male/female ratio, inhibition level, degradation level, etc. These quality checks can help you identify potential issues or errors in your samples and correct them before performing STR analysis.
The QuantStudio 5 HID system can be used for various human identification applications that require real-time PCR. The system can help you achieve accurate, sensitive, robust, and fast results that can help you solve cases, identify individuals, and establish relationships.
Benefits and Drawbacks
The QuantStudio 5 HID system has some benefits and drawbacks based on its features and performance. Some of the benefits and drawbacks are:
Benefits:
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- The QuantStudio 5 HID system has a streamlined design that saves bench space, simplifies operation, and enhances user experience. The system has a small footprint, front-facing motorized block, touch screen interface, USB connectivity, and onboard memory.
- The QuantStudio 5 HID system has interactive software that helps users design, run, and analyze real-time PCR experiments with ease and flexibility. The software includes the QuantStudio 5 DAS and the HID RAS that support various analysis methods, reports, and graphs. The software also allows users to access their data from anywhere using a web browser or a mobile app.
- The QuantStudio 5 HID system delivers reliable performance that ensures consistent results, long-term durability, data storage, and quality assurance. The system has a bright white LED source, less frequent maintenance, self-cleaning and self-diagnostic features, and HID validation.
- The QuantStudio 5 HID system incorporates an innovative technology that enables future quantification technology, optimizes reaction conditions, and customizes thermal profiles. The system has a multiplexing capability for up to six dyes per well, precise temperature control with Peltier-controlled blocks, and VeriFlex zones.
Drawbacks:
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- The QuantStudio 5 HID system may require calibration with different dyes depending on the assay used. The system is factory-calibrated with FAM, SYBR Green I, VIC, NED, ABY, JUN, Mustang Purple, TAMRA, Cy5, and ROX dyes. However, in order to run the HID assay, HID ABY and HID JUN dyes will be delivered with each instrument and will require calibration upon installation.
- The QuantStudio 5 HID system may not be compatible with some real-time PCR kits or consumables from other manufacturers. The system is optimized and validated for human identification applications using Applied Biosystems quantification kits and Y-STR kits. The system also supports 96-well 0.2 mL or 0.1 mL format or 384-well format for consumables.
- The QuantStudio 5 HID system may have some limitations in terms of throughput or scalability. The system can run up to 96 samples per run in the 96-well format or up to 384 samples per run in the 384-well format. However, the system does not have an option for automation or integration with other instruments or platforms.
The QuantStudio 5 HID system has some benefits and drawbacks based on its features and performance. Users should weigh these benefits and drawbacks according to their needs and preferences when considering the QuantStudio 5 HID system for their human identification applications.
Applied BioSystems Quantstudio Evolution
The QuantStudio 5 HID system is the latest addition to the Applied Biosystems HID real-time PCR family of products, which includes the previous models such as the 7500 Real-Time PCR System and the 7500 Fast Real-Time PCR System.
The QuantStudio HID system has evolved from previous models to provide improvements, address issues, or otherwise help users in making a purchase decision.
Some of the improvements and advantages of the QuantStudio 5 HID system are:
- The QuantStudio 5 HID system has a smaller footprint, front-facing motorized block, touch screen interface, USB connectivity, and onboard memory, which save bench space, simplify operation, and enhance user experience.
- The QuantStudio 5 HID system has a multiplexing capability for up to six dyes per well, which enables future quantification technology and allows the detection of more targets in a single reaction.
- The QuantStudio 5 HID system has precise temperature control with Peltier-controlled blocks and VeriFlex zones that allow customization of thermal profiles and optimization of reaction conditions.
- The QuantStudio 5 HID system has interactive software that includes the QuantStudio 5 DAS and the HID RAS that enable easy design, running, and analysis of real-time PCR experiments with various analysis methods and reports.
- The QuantStudio 5 HID system has a bright white LED source that provides uniform illumination and reduces optical variation. The LED source also has a long lifespan that reduces the need for replacement or calibration.
- The QuantStudio 5 HID system has less frequent maintenance and self-cleaning and self-diagnostic features that ensure consistent results, long-term durability, and quality assurance.
- The QuantStudio 5 HID system is compatible with various quantification kits and Y-STR kits from Applied Biosystems that offer high sensitivity, robustness, discrimination capacity, and multiplexing capability for human identification applications.
The QuantStudio 5 HID system is a reliable and affordable real-time PCR system for human identification applications that have evolved from previous models to provide improvements, address issues, or otherwise help users in making a purchase decision.
Similar Alternatives to Applied BioSystems QuantStudio 5
The QuantStudio 5 HID real-time PCR system for human identification has certain similarities and differences with other Applied Biosystems and other real-time PCR systems.
Some of the similarities and differences are:
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Similar Design:
The QuantStudio 5 HID system is similar to the QuantStudio 3 and 5 Real-Time PCR Systems but optimized for human identification applications. It includes HID RAS software for analysis and reporting.
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Features:
The QuantStudio 5 HID system has a similar footprint, block format, reaction volume, and run time as the 7500 Real-Time PCR System and the 7500 Fast Real-Time PCR System. However, the QuantStudio 5 HID system has a front-facing motorized block, touch screen interface, USB connectivity, on-board memory, bright white LED source, less frequent maintenance, multiplexing capability for up to six dyes per well, precise temperature control with Peltier-controlled blocks and VeriFlex zones, and interactive software that make it more user-friendly, reliable, and innovative than the 7500 systems.
- The QuantStudio 5 HID system differs from other real-time PCR systems like Bio-Rad CFX96 Touch or Qiagen Rotor-Gene Q. It has a smaller footprint, front-facing motorized block, touch screen, USB connectivity, on-board memory, bright white LED source, less frequent maintenance, multiplexing capability for up to six dyes per well, precise temperature control with Peltier-controlled blocks and VeriFlex zones, and interactive software that includes the QuantStudio 5 DAS and the HID RAS for easy design, run, and analysis of real-time PCR experiments with various analysis methods and reports. These features make the QuantStudio 5 HID system more suitable for human identification applications than other real-time PCR systems.
Conclusion
The QuantStudio 5 HID system is a reliable and affordable real-time PCR system for human identification applications. Its simplified design, engaging software, dependable performance, and cutting-edge technology make it the perfect option for human identification laboratories. Numerous human identification applications, including forensic databases, forensic casework, confirmatory screening, sexual assault sample workflow, quality and quantity sample assessment, etc., can be performed using the system. You can easily and confidently use the system to accomplish your human identification goals.
If you are interested in learning more about the QuantStudio 5 HID system or want to request a quote or a demo, please visit our website or contact our sales representative. We would be happy to assist you with any questions or inquiries you may have.
Frequently Asked Questions
Here are some frequently asked questions about the QuantStudio 5 HID system and their answers:
What are the main differences between the QuantStudio 5 HID system and the previous Y-STR systems from Applied Biosystems?
The main differences are:
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- The QuantStudio 5 HID system has a smaller footprint, front-facing motorized block, touch screen interface, USB connectivity, and on-board memory.
- The QuantStudio 5 HID system can multiplex up to six dyes per well, enabling future quantification technology.
- Peltier-controlled blocks and VeriFlex zones offer accurate temperature control in the QuantStudio 5 HID system.
- The QuantStudio 5 HID system’s interactive software, including the DAS and RAS, simplifies real-time PCR experiment planning, execution, and analysis.
- The QuantStudio 5 HID system has a bright white LED source that provides uniform illumination and reduces optical variation.
- The QuantStudio 5 HID system has less frequent maintenance and self-cleaning and self-diagnostic features.
What are the advantages of using the Yfiler Plus PCR Amplification Kit with the QuantStudio 5 HID system?
The advantages are:
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- The Yfiler Plus PCR Amplification Kit amplifies 27 Y-STR loci in one reaction, including seven rapidly-mutating (RM) STRs that improve paternal lineage separation and male discrimination.
- The Yfiler Plus PCR Amplification Kit’s excellent sensitivity and resilience allow allele recovery from degraded or hindered material.
- The QuantStudio 5 HID system’s multiplexing capabilities can detect up to six dyes per well with the Yfiler Plus PCR Amplification Kit.
- The Yfiler Plus PCR Amplification Kit works with HID RAS software to analyze and interpret Y-STR results.
How can I calibrate the QuantStudio 5 HID system with different dyes?
The QuantStudio 5 HID system is factory-calibrated with FAM, SYBR Green I, VIC, NED, ABY, JUN, Mustang Purple, TAMRA, Cy5, and ROX dyes. In order to run the HID assay, HID ABY (Cat. No. 4461591) and HID JUN (Cat. No. 4461593) dyes will be delivered with each instrument and will require calibration upon installation. You can use the touch screen interface or the DAS software to perform the calibration with different dyes as needed.
How can I access my data from the QuantStudio 5 HID system?
The DAS software on the instrument or the touch screen interface both provides access to your data. For backup or additional analysis, you can also export your data to a PC or USB drive. With the HID RAS software, you can use a web browser to access your data from any location.
How can I perform quality checks on my samples using the QuantStudio 5 HID system?
HID RAS can calculate the male/female ratio, inhibition level, degradation level, etc. for sample quality checks. These quality tests can help you find and fix sample issues before STR analysis.
What are the specifications and requirements of the QuantStudio 5 HID system?
The specifications and requirements of the QuantStudio 5 HID system are:
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- Dimensions and weight: 27 x 50 x 40 cm (W x D x H), <26 kg
- Sample capacity (wells): 96-well 0.2 mL block
- Reaction volume: 96-well: 10–100 μL for 0.2 mL block
- Maximum ramp rate: 6.5°C/sec
- Average sample ramp rate: 3.66°C/sec
- Temperature uniformity: 0.4°C
- VeriFlex Blocks: 96-well: 6 independent temperature zones
- Heating/cooling method: Peltier
- Run time: ~70 min for Quantifiler Trio and HP kits; ~100 min for Quantifiler Duo and Human kits
- Calibration: Factory-calibrated*
- Onboard memory: 10 GB, which translates to approximately 2,000–5,000 run files
- Electrical approvals: IEC, CE
What are the advantages of using VeriFlex zones with the QuantStudio 5 HID system?
The advantages of using VeriFlex zones with the QuantStudio 5 HID system are:
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- You can run several experiments or optimise different parameters in the same block by creating different temperature zones.
- You have more control over thermal profiles by setting temperatures within ±15°C of the blocking temperature.
- VeriFlex zones can detect sequence changes in DNA samples using high resolution melting (HRM) analysis.
How can I perform multiplexing with the QuantStudio 5 HID system?
The QuantStudio 5 HID system can multiplex up to six dyes per well with Quantifiler Trio and HP DNA Quantification Kits. These kits evaluate total human DNA, male DNA, IPC, DI, and II in one reaction. The six-dye Yfiler Plus PCR Amplification Kit amplifies 27 Y-STR loci in one reaction.
How can I perform direct amplification with the QuantStudio 5 HID system?
You can directly amplify single-source samples with the QuantStudio 5 HID system using the Yfiler Plus PCR Amplification Kit. The kit can amplify DNA from blood and buccal samples on treated paper, blood on untreated paper treated with Prep-n-Go Buffer, or swabs treated with Prep-n-Go Buffer. No sample purification is needed.
