QIAcuity EG PCR Kit

For use with the QIAcuity digital PCR instruments


  • For dye-based digital PCR reactions using EvaGreen
  • 3x concentrated Master Mix for loading more sample
  • Optimized for microfluidic use in QIAcuity Nanoplates
  • REACH compliant

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QIAcuity EG PCR Kit (1 ml)

Cat. No. / ID: 250111

1 ml 3x concentrated QIAcuity EvaGreen Mastermix, 2 x 1.9 ml Water
1 ml
5 ml
25 ml
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The QIAcuity EG PCR Kit is intended for molecular biology applications. This product is 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

Product Details

The QIAcuity EG PCR Kit contains a 3x concentrated, ready-to-use Master Mix optimized for microfluidic use in the QIAcuity Nanoplates. The kit improves the specificity and efficiency of dye-based digital PCR to provide accurate quantitation analysis. An intercalating dye EvaGreen binds to double-stranded DNA and enhances the quantitative accuracy of gDNA or cDNA measurements on the QIAcuity dPCR instruments.


The kit works in conjunction with the QIAcuity Digital PCR System and the QIAcuity Nanoplates.


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Superior performance
The QIAcuity Master Mixes for EvaGreen-based detection use the latest versions of QIAGEN’s high-quality DNA Polymerase. The unique combination of QIAGEN's proprietary and well-proven buffer technology optimized for the Nanoplate microfluidic along with the new QuantiNova DNA Polymerase delivers highly consistent results in terms of sensitivity, reproducibility and efficiency.


Dye-based detection with EvaGreen
The special master mix in the QIAcuity EG PCR Kit enables accurate double-stranded DNA target amplification and quantification. It includes an optimized reference dye needed for dPCR analysis and counting analyzable partitions in Nanoplates. Moreover, EvaGreen provides a higher fluorescence signal than SYBR Green at the same concentrations and delivers maximum amplification efficiency, specificity and sensitivity in dPCR. 


Reaction stability of up to 100 hours
The QIAcutiy PCR mixes can be stored at 30°C for up to 100 hours without impairing the performance of subsequent reactions. The outstanding stability, even after extended storage at room temperature without the use of any cooling agent, makes the QIAcuity EG PCR Kit ideal for high-throughput reaction setup and plate-stack handling.


The QIAcuity EG PCR Kit delivers cDNA or gDNA analysis with the highest specificity because of a novel, antibody-mediated, hot-start mechanism. At low temperatures, the QuantiNova DNA Polymerase is kept in an inactive state by the QuantiNova Antibody and a novel additive, QuantiNova Guard, that stabilizes the complex. This improves the stringency of the hot-start and prevents extension of nonspecifically annealed primers and primer–dimers. Within 2 minutes of raising the temperature to 95°C, the QuantiNova Antibody and QuantiNova Guard are denatured, and the QuantiNova DNA Polymerase is activated, enabling the PCR amplification.

The principle of the dPCR reaction in the nanoplates is described here.


Just like in qPCR experiments, sample preparation includes the transfer of master mix, probes and primers to a 96- or 24-well nanoplate, followed by the addition of samples. The system integrates partitioning, thermocycling and imaging into a single fully automated instrument that takes users from the sample to result in under 2 hours. One can perform analysis on the Suite Software, providing the concentration in copies per microliter of your target sequence as well as for quality control such as positive samples or NTC. This analysis can also be extended to remote computers within the same local area network (LAN).


The QIAcuity EG PCR Kit, in combination with the QIAcuity Digital PCR System and the QIAcuity Nanoplates, enable quantitative analysis of cDNA targets and gDNA for use in applications, including:

  • Rare mutation detection
  • Copy number variation analysis
  • Gene expression analysis
  • Pathogen detection
  • Genotyping
  • miRNA research

Supporting data and figures


Operating Software (6)
QIAcuity Software Suite
Version 1.1.3
Version 2.0

The Volume Precision Factor (VPF) offers a unique feature to secure precision of concentration results obtained from a QIAcuity dPCR run. 
In general, Nanoplates provide partitions of fixed sizes that enable a very precise way of sample concentration calculation. Potential variation of partition sizes in Nanoplate batches, caused by different microstructure molding forms, can be addressed by applying the batch specific VPF. Furthermore, the VPF includes well-specific volume information and therefore further increases precision of concentration calculation in each well of the Nanoplates.

After downloading and updating the VPF file within the QIAcuity Software Suite, the VPF is applied automatically to the analysis of a corresponding Nanoplate batch. The VPF file includes information from all available microstructure molding forms and connected Nanoplate batches. It will be stored on the PC where the QIAcuity Software Suite is installed. 

Required QIAcuity Software Suite version: Version 1.2 or higher.
QIAcuity Control Software
Version 1.0

The QIAcuity Control Software is an integral part of the QIAcuity instrument. It offers a GUI (graphical user interface) for basic functionalities such as plate setup, changing the order of plates to be processed, and monitoring the status of runs in real time. After a run is completed, the data are stored on the instrument’s memory and are sent to the connected QIAcuity Software Suite for analysis.

This new version of the control software includes system stability and performance reliability improvements. We highly recommend to update the control software of your QIAcuity instrument.

Note: The installation requires approximately 90 minutes.
QIAcuity Software Suite

Version 1.2

The QIAcuity Software Suite 1.2 is designed to be installed on a Windows PC that is connected to one or more QIAcuity instruments. The QIAcuity Software Suite enables the user to set up plates, analyze results, and monitor the status of runs in real time. For this configuration, the QIAcuity instrument needs to be connected to a network through Ethernet. Alternatively, a direct cable connection between the QIAcuity and the notebook where the QIAcuity Software Suite is running needs to be established. When connected to a network, up to 10 users may access the QIAcuity Software Suite via a browser installed on the client PC (Windows or Mac).

The following browsers are supported in the QIAcuity Software Suite:

-Mozilla Firefox (version 64.0.2 or higher)
-Microsoft Edge (version 44.17763.1.0 or higher)
-Google Chrome (version 71.0.3578.98 or higher)

The new QIAcuity Software Suite 1.2 offers a functionality that enables users of the QIAcuity Software 1.1.3 to upgrade to the new version while keeping the library of previously stored plate runs.

Note: If you have exported plates from QIAcuity Software Suite 1.1.3 that you would like to import and use in QIAcuity Software Suite 1.2, you will need to import the plates before upgrading from version 1.1.3 to version 1.2. You may then export the plates again. Future software version starting from QIAcuity Software Suite 2.0 will facilitate import of plates from previous QIAcuity Software Suite versions.

The new improvements are as follows:

-Support for the Nanoplate 8.5k 24-well
-Hyperwell functionality to combine several wells to one combined well for analysis
-Automated plate archiving functionality
-Functionality to show the number of single/double positives in 2D scatterplots
-VPF (Volume Precision Factor) to further improve concentration calculation (see related resources)
-Additional improvements for stabilization and troubleshooting

For Version 1.2
Instrument User Manuals (2)
Webinars (5)
Limitations of conventional PCR and qPCR when dealing with difficult, low-volume samples and complex mixtures with high background of competitive molecules and inhibitors have posed frequent challenges for researchers and clinicians in their routine work. With the new generation of PCR technologies, digital PCR has opened doors for diverse applications, and researchers are learning to ask questions only digital PCR can answer. Join QIAGEN's webinar on how digital PCR can help take your research applications through and beyond those challenges.
This presentation will introduce dPCR, discuss its advantages, and outline how the approach might be used to improve measurement in areas like clinical diagnosis, alone or in conjunction with other methods.
In this expert webinar, Dr. Kubista will share with you the experience he and his team have gathered at the TATAA Biocenter, developing applications and providing services using digital PCR for nearly 12 years. They have experienced all the problems common to dPCR analytical workflows and developed robust standard operating procedures to minimize the risk of error and maximize robustness and repeatability, and developed various controls to test the performance and validate the methods. He will also discuss dPCR assay design and validation and then focus on strategies for copy number determination and rare mutation detection.
The QIAGEN digital PCR technology and its expanded capability will not only transform the portfolio of conventional qPCR applications but also provide a more rapid, accurate, and sensitive method for finding answers to difficult biological questions. 
As the digital PCR technology evolves and becomes more accessible and affordable, the transition from qPCR and adoption of dPCR will hopefully no longer remain a challenge. Experts share insights in an upcoming webinar about the fully integrated, rapid, and highly flexible digital PCR portfolio from QIAGEN. 
Kit Handbooks (1)
For highly sensitive detection of miRNA using EvaGreen


Can I use the QIAcuity Nanoplate in more than one runs?

The plate is designed for a single use run. For example, even if only 30 samples are loaded into the 96-well plate, a whole plate will be sealed by the roller. It can't be unsealed and used for another run. The QIAcuity Software won’t allow to set up a separate experiment for the same nanoplate to avoid that previously processed plates being not partitioned a second time.

Can I prepare a dPCR reaction directly in QIAcuity Nanoplate?

A standard PCR plate is required to set up dPCR reaction before transferring it to the nanoplate to ensure a proper mixing of the reaction mix before partitioning. 

Where can I find assays specifically designed for QIAcuity?

QIAGEN dPCR assays (such as dPCR LNA Mutation Assays) can be found on https://geneglobe.qiagen.com/.

Is it possible to change voltage set-up from 110V to 230V on the QIAcuity instruments?

This is not needed. The QIAcuity is equipped with a flexible power supply technology and operates within a range of 100–240V AC, 50/60 Hz, 1500 VA (max). 

What is the impact of not applying the latest VPF? Can I reanalyze previously obtained results after installing the latest VPF?

If you had run a nanoplate for which the installed VPF misses the specific factor, the software will notify you. If you then analyze without the specific VPF, the impact depends on the variation of the partition volume of the new Nanoplate batch compared to the latest. Typically this variation is ±6–7% (approx. 5% CV over the entire plate). The analysis may be repeated after updating the VPF file. After installing the latest VPF and re-analysis of the run, a copy of the plate is generated in the QIAcuity Software Suite including the new results. 

Can I use a custom master mix instead of a QIAGEN master mix?

QIAGEN master mixes are optimized for nanoplate microfluidics and are recommended to be used with our dPCR system. They also include an optimized reference dye required for proper analysis.

Can I optimize a dPCR assay on the QIAcuity using gradient temperature?

QIAcuity does not support temperature gradient in PCR cycling profile. However, optimization of a dPCR assay can be done by qPCR on a gradient cycler using the dPCR master mix and then transferred from qPCR to dPCR. 

Which sample preparation method is recommended for QIAcuity dPCR?

QIAGEN offers a complete range of nucleic acid purification systems. These include QIAprep kits for purification of plasmid DNA, QIAamp, and DNeasy kits for purification of genomic DNA, RNeasy kits for purification of total RNA, and the PAXgene Blood RNA System for stabilization and purification of RNA from blood. Phenol and other contaminants can be efficiently removed from crude RNA preps using the RNeasy MinElute Cleanup Kit to clean up and concentrate RNA for sensitive assays. Details about QIAGEN kits for nucleic acid purification can be found at www.qiagen.com.

Is it necessary to reanalyze a plate with VPF (Volume Precision Factor) that was already processed using a QIAcuity instrument that was purchased in 2020?

If you had analyzed your nanoplates without VFP, the impact depends on the variation of the partition volume of the new nanoplate batch compared to the latest. The VPF reduces the CV from approximately 5% to 2%. We recommend to reanalyze results in case the data originated from different wells (e.g., copy number variations or gene expression data sets for which the reference sample was measured in a different well). Results obtained across different plates should also be r-analyzed. A reanalysis is not required for assay data that were analyzed within the same well (e.g., mutation rate determination using two channels within the same well).

Is a standard curve needed in dPCR?

In dPCR we measure the absolute concentration of targets at endpoint reaction. Concentrations of unknowns can be determined based on dPCR results observed (number of negatives, number of positives, and partition volume analyzed).

When and how often do I need a new VPF (Volume Precision Factor)?

 In general, nanoplates provide partitions of fixed sizes that enable a very precise way of sample concentration calculation. If a new molding form is used for nanoplate manufacturing, potential variation of partition sizes can be addressed by applying the molding form-specific VPF. Thus, each time a new molding form is used, a new VPF is created and made available. Currently, the VPF is updated once every 3–6 months.

What is the purpose of the reference channel in QIAcuity?

The fluorescent signal in the reference channel is measured to determine the number of valid partitions in a well. In addition, differences in the signal intensities between partitions are normalized and the fluorescent signals in the target channels are corrected accordingly.

What is the VPF (Volume Precision Factor)?

The VPF provides a set of well-specific and molding form-specific factors used to specify the exact reaction volume of a nanoplate, thus increasing the concentration calculation of each well. 

Can I see on the QIAcuity Software Suite report file if the VPF (Volume Precision Factor) has been used or not?

Yes, the report includes a notification if the matching VPF was missing and, therefore, not applied to the analysis. If the matching VPF was applied there is no notification on the report. 

What are the storage conditions and expiry date of QIAcuity consumables?

The QIAcuity Probe PCR Kit should be stored immediately upon receipt at –30 to –15°C in a constant-temperature freezer and protected from light. The QIAcuity Probe PCR master mix can also be stored protected from light at 2–8°C. Components are stable for 12 months, unless otherwise indicated on the label. 
The QIAcuity EG PCR Kit should be stored immediately upon receipt at –30 to –15°C in a constant-temperature freezer and protected from light. The QIAcuity EG PCR master mix can also be stored protected from light at 2–8°C. Components are stable for 6 months, unless otherwise indicated on the label. 
The QIAcuity Nanoplates does not have expiry date and are stable for at least 1 year when stored at RT. 

What is the input volume for QIAcuity Nanoplate well?

Nanoplate 26K 24-well: 40 μl 
Nanoplate 8.5K 24-well:12 μl 
Nanoplate 8.5K 96-well: 12 μl 

How to prepare DNA prior to dPCR?

All DNA samples used in reaction mixes should show similar quality and quantity, which can easily be assessed using UV spectrophotometry. DNA samples with an average length of ≥20 kb (e.g., genomic DNA purified via spin column with silica membrane) should be fragmented by restriction digestion before partitioning. Enzymatic fragmentation of larger DNA ensures an even distribution of template throughout the QIAcuity Nanoplate, which in turn leads to an accurate and precise quantification.

Can I see error codes on the instrument touchscreen?

The instrument software GUI shows error codes including a description and information how to resolve the error. The instrument touchscreen shows an alarm icon in the upper right corner that turns red in case of an instrument failure. Accessing the System Status in the Tool tab allows users to clear errors. Rebooting of the instrument is required to complete the removal of the error.  Please do not skip this step. You may always contact QIAGEN Technical Services in case of any question. 

Can I use an empty nanoplate without seal to perform a dry run?

The QIAcuity instrument software does not allow to read and process a plate without seal. If you would like to perform a dry run please use sealed plate and set up this plate in the QIAcuity Software Suite. 

Can I use the dPCR master mix in a qPCR cycler for optimization purposes?

dPCR master mix can be used in qPCR to optimize sample concentration and/or primer/probe concentration prior to assay transfer from qPCR to dPCR. 

What is the scope of a regular maintenance of the QIAcuity?

The user manual contains instructions on how to perform a regular cleaning and decontamination, and how to replace air filters on the QIAcuity instruments. A regular maintenance reduces the dust in the instrument and therefore minimizes the presence of dust particles on the nanoplate, which might interfere with the plate analysis.

How can I get the latest VPF (Volume Precision Factor)?

The up-to-date VPF can be downloaded in the resource section of the QIAcuity product webpage. The VPF  is compatible with the QIAcuity Software Suite 1.2 and higher. The latest VPF file contains all factors for all existing nanoplate batches. If a nanoplate from a new nanoplate batch is run and the latest version of the VPF was not installed, the software will recognize this and will give a message to install the latest VPF file. Please note that Applying the VPF file cannot be reversed.

Does an incorrect handling of a plate might affect the results?

The QIAcuity reads emitted fluorescence from the bottom of the plate, which is covered with a foil. For best results, keep the foil clean and avoid damages such as scratches. Also, keep the barcode on the side of the plate clean and intact. Ensure that you wear gloves when working with a plate and do not apply force to it. For a safe handling of the plate, please  place the plate into a nanoplate tray.

Can the QIAcuity One 2plex be upgrade to the QIAcuity One 5plex?

No. The QIAcuity platform introduces four variations: QIAcuity One 2plex, QIAcuity One 5plex, QIAcuity Four (5plex), and QIAcuity Eight (5plex). All of them have fix channel combination. 

How are results stored within the QIAcuity Software Suite?

Results are stored as part of the plates within the QIAcuity Software Suite. In version 1.1.2, plates can be exported to another file location, for example, an external HDD, and imported again if needed. From version 1.2 onwards, plates can also be archived automatically. To do so, an archive destination has to be defined. Additional information can be found in the user manual. 

How can I upgrade the software on the instrument (CSW) and the QIAcuity Software Suite?

Both software are designed to be upgraded by users. The user manual includes instructions on how to perform the upgrades; instructions for the QIAcuity Suite Software upgrade can be found at page 38 and instructions for the CSW upgrade on page 67 (QIAcuity User Manual, 03/2021).