Cell and gene therapy research
Measure common viral titer backbones, custom genes of interest, or the presence of residual DNA without the use of a standard curve for faster, simpler, more precise quantification.
Improve detection of mutations and quantification of low allele fractions in challenging samples such as liquid biopsy with high specificity, precision and accuracy of dPCR.
Accurately measure viral load and pathogens or other biomarkers in environmental samples such as wastewater by absolute quantification with improved performance compared to qPCR in the face of common PCR inhibitors.
Inherited disease research
Accurate and precise quantification of rare mutations associated with human diseases along with exceptional discrimination of CNVs with high copy number.
Detect genome editing events in a fast and reliable way, especially those created using nucleases that cause double-stranded breaks, such as CRISPR-Cas9.
Digital PCR is a specialized approach to nucleic acid detection and quantification that estimates absolute numbers of molecules through statistical methods. The digitization process distributes the PCR mix across thousands of microreactions so that each microreaction will effectively either contain one, zero, or just a handful of the target nucleic acid molecules. In effect, the original reaction is turned into many yes-or-no reactions. After counting the positive microreactions, simple statistics can be used to then determine the “absolute” quantity of the target molecule rather than a quantity estimated by comparing to a standard of known concentration.
As the name suggests, real-time PCR measures PCR amplification as it occurs. It focuses on the exponential phase because it provides the most precise and accurate data for quantitation. By comparing the Ct values of samples of unknown concentration with a series of standards, the amount of template DNA in an unknown reaction can be accurately determined.
Digital PCR works by digitizing a sample into many individual real-time PCR reactions; some portion of these microreactions contain the target molecule (positive) while others do not (negative). Following PCR analysis, the fraction of negative answers is used to generate an absolute answer for the exact number of target molecules in the sample, without the need for Ct values or reference to standards.
dPCR allows you to simply count the number of microchambers that have a specific or set of specific target molecules. This provides a mechanism for absolute quantification, which lets you determine the number of molecules in your sample without the need for standard curve as would be typical in a qPCR reaction. This standard curve–free absolute quantification is especially beneficial for laboratories in need of highly precise and consistent quantification methods. Since dPCR also allows you to identify handfuls of rare molecules in an overwhelming number of normal or background molecules, it is excellent for applications such as detecting rare oncogenic mutations in circulating free DNA. Multiplexing enables you to perform absolute quantification to several target molecules in parallel.
Additionally, dPCR takes an endpoint measurement in order to determine concentration and is overall less sensitive to some common inhibitors that can interfere with qPCR reaction kinetics and efficiency. This improved performance in the face of inhibitors is an important attribute for research labs working with environmental or precious samples.
dPCR is well suited to performing rare allele detection, measurement of copy number variation, viral titer measurement, quantification of next-generation sequencing libraries, and detection of rare targets from environmental samples such as wastewater. Specific applications available for absolute quantification include:
The QuantStudio Absolute Q Digital PCR System is a plate-based dPCR platform powered by proprietary microfluidic array plate (MAP) technology that enables all the necessary steps for dPCR—compartmentalizing, thermal cycling, and data acquisition—to be conducted on a single instrument. The dPCR workflow is identical to the qPCR workflow you are familiar with to improve ease of use, minimize hands on steps, and maximize consistency. Absolute Q dPCR assays, which can be used for liquid biopsy research and viral titer measurement, can also be customized to answer unique questions. The assays are based on the same gold-standard TaqMan chemistry used in real-time PCR and cited in more than 200K research publications.
Whether you are new to digital PCR or an experienced user looking to learn new tricks, we have a library of educational material to help you.
Learn how to quickly get started using the QuantStudio Absolute Q dPCR System
View quick reference guide ›
In this presentation, learn about the latest innovations to enable absolute molecular quantification. The automation compatible QuantStudio Absolute Q Digital PCR system will be highlighted, showcasing high throughput consistent data without the need for standard curves.
Clarence Lee, Ph.D.
Sr. Product Manager, Digital PCR, Thermo Fisher Scientific
Our panel of key opinion leaders in the liquid biopsy space will discuss the impact of genomics technologies such as dPCR and NGS on the current state and future outlook of liquid biopsy clinical research and cancer management.
In the featured videos below, see the fast, simple, and robust QuantStudio Absolute Q Digital PCR System in action.
Learn why the QuantStudio Absolute Q dPCR System is the number one solution for simple, fast, and reliable digital PCR.
Getting started with the QuantStudio Absolute Q dPCR System is simple and takes under five minutes to prepare a MAP16 digital PCR plate for a run.
Learn why our customers chose powerfully simple digital PCR to meet their unique research needs.
A digital PCR-based protocol to detect and quantify RNA editing events at hotspots
Development and validation of a 4-color multiplexing spinal muscular atrophy (SMA) genotyping assay on a novel integrated digital PCR instrument
SARS-CoV-2 RNAemia predicts clinical deterioration and extrapulmonary complications from COVID-19
Precision cancer monitoring using a novel, fully integrated, microfluidic array partitioning digital PCR platform
For Research Use Only. Not for use in diagnostic procedures.