Digital Assays Part I: Partitioning Statistics and Digital PCR
11 Department of Electrical and Computer Engineering, and Department of Biomedical Engineering, Wayne State University, Detroit, MI, USA.
SLAS Technology
|April 28, 2017
Summary
Digital assays offer precise quantification of biological entities by partitioning samples. This review focuses on digital PCR (dPCR) applications for nucleic acid analysis, highlighting its advantages over quantitative real-time PCR.
Area of Science:
- Biotechnology
- Molecular Biology
- Genomics
Background:
- Digital assays partition samples into discrete containers for precise biological entity counting.
- These assays enhance quantification of nucleic acids, proteins, and single-cell properties.
Purpose of the Study:
- To review the principles, benefits, and applications of digital assays, with a focus on digital PCR (dPCR).
- To compare dPCR with quantitative real-time PCR (qPCR) and discuss its utility in various scientific workflows.
Main Methods:
- Review of partitioning statistics (Poisson distribution) and sources of error in digital assays.
- Analysis of five commercial chamber (cdPCR) and droplet (ddPCR) digital PCR instruments.
- Comparison of dPCR capabilities (absolute quantitation, rare target detection) against qPCR (dynamic range, throughput).
Main Results:
- Digital PCR provides absolute quantitation, high precision, and superior detection of rare targets compared to qPCR.
- Commercial instruments offer diverse platforms for implementing dPCR in nucleic acid quantification.
- dPCR demonstrates broad applicability in areas like copy number variation, ctDNA, viral load, and NGS library prep.
Conclusions:
- Digital assays, particularly dPCR, represent a significant advancement for precise biological quantification.
- dPCR offers distinct advantages over qPCR for specific applications, enabling new research possibilities.
- This review provides a foundational understanding for scientists integrating digital assays into their research.
