Choosing dPCR vs. qPCR – Explore the benefits and see if it’s right for your research goals
When comparing dPCR vs. qPCR technologies, the key difference is precision power. While both offer highly sensitive and reliable nucleic acid detection and quantification, the key difference between the two technologies can be best described using an analog versus digital radio analogy, says Dr. Jim Huggett, Principal Scientist, National Measurement Laboratory. "With an analog radio, the dial must first be fine-tuned to get the desired station with the least interference. Still, the quality depends on reception and the signal is subject to interference from static. This is qPCR. It is reliable but requires optimization to get a good result, and even then, you must contend with background noise. With digital radio, you simply call up the station and it is either there, with a clear signal, or not. The latter is like dPCR, which provides precise, binary results. It literally counts the presence or absence of DNA molecules. The clarity of results combined with a low error rate makes for an incredibly high level of precision. Digital PCR is well-suited to measure smaller quantitative differences."
Compare and contrast: qPCR vs. dPCR
| Real-time PCR/ qPCR | Digital PCR |
|---|---|
| Quantitative, relative or absolute but standard curves or reference samples needed |
Quantitative, absolute and no standards or references needed |
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Bulk PCR
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Sample partitioning
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| Measures PCR amplification at each cycle | Measures at the end of the PCR cycles |
| Detects mutation rate at >1% | Detects mutation rate at ≥ 0.1% (high signal-to-noise ratio) |
| Well-established protocols | Higher precision for higher reproducibility across laboratories |
When to use dPCR vs. qPCR?
When it comes to molecular biology and genomics research involving nucleic acid quantification, scientists often find themselves standing at the crossroads. Which quantification technique to choose to achieve research goals efficiently – the more precise and robust digital PCR (dPCR) or the more standardized and familiar quantitative real-time PCR (qPCR). Both technologies have similarities, but also their benefits and limitations that make the choice application-dependent.
The application grid indicates the suitability level of each technology for some of the common applications.
Droplet digital PCR (ddPCR), one of the earliest forms of digital PCR, could already offer advantages over qPCR for most of the applications above. In ddPCR vs qPCR, qPCR is suitable for applications that require a broad dynamic range, whereas ddPCR is for those applications that require higher precision or analysis of fractional abundance.
The evolution from ddPCR to nanoplate dPCR has widened the scope of this technology to include more applications. The nanoplate dPCR workflow is substantially faster thanks to the simultaneous reading of all sample partitions, front-end automation, and an easy qPCR-like plate set-up. This added speed makes it suitable for screening and high-throughput applications without compromising precision, accuracy and sensitivity.
Researchers describe the benefits of dPCR over qPCR – See examples of a few studies that used dPCR to overcome the limitations of qPCR.
Transitioning from qPCR to dPCR – The QIAcuity Nanoplate Digital PCR System makes it accessible and more affordable than ever to bring the power of dPCR to your lab.
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Transforming the PCR experience
Imagine keeping the familiarity and ease of qPCR but gaining the higher sensitivity and precision of digital PCR, without prolonging the time to scientific insight. The QIAcuity, QIAGEN's fully integrated nanoplate-based digital PCR system, has been designed with your research needs and the limitations of the currently available methods in mind. -
How does QIAGEN Nanoplate dPCR work?
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 remote analysis on the Software Suite, providing the concentration in copies per microliter of your target sequence as well as for quality control such as positive samples or NTC.
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Unveiling the magic inside
Reimagining familiarity, ease of use and finer resolution, we have developed these nanoplates with superior partitioning, meaning unparalleled precision and sensitivity gain for your assays. -
No droplets. No crystals. No chips.
The key advantages of performing digital PCR in nanoplates include but are not limited to:
- User-friendly, familiar plates are easy to pipet just like for qPCR
- Fixed partitions prevent variation in size and coalescence
- Sealed nanoplates prevent well to well contamination
- Faster readout possible due to simultaneous reading of all partitions of a sample
- Integrated quality control, meaning one can look at the plate image for single fluorescent signals from the individual positive partition counts
- Plates are amenable to front-end automation -
Simplifying going digital
It's time to simplify the transition from qPCR and prepare your critical research applications to benefit from digital PCR in more ways than one. Yes, in just 3 simple steps you can have the accurate result you want in under 2 hours: pipette and load, let the experiment run, analyze results. Moreover, cost, scalability and throughput are no longer a bottleneck when considering that switch.
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Transforming the PCR experience
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How does QIAGEN Nanoplate dPCR work?
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Unveiling the magic inside
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No droplets. No crystals. No chips.
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Simplifying going digital