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PCR01:32

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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Updated: Mar 10, 2026

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays
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Digital PCR: A brief history.

Alexander A Morley1

  • 1Department of Haematology and Genetic Pathology, Flinders University and Medical Centre, Bedford Park, SA 5042, Australia.

Biomolecular Detection and Quantification
|December 7, 2016
PubMed
Summary

Digital PCR, a method for quantifying targets, has evolved significantly since its inception. Recent advancements in instrumentation and chemistry have revitalized digital PCR, making it more accessible and practical for researchers.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Digital PCR (dPCR) was first described as "limiting dilution PCR" in 1990-1991 and later as "digital PCR" in 1999.
  • Its initial adoption was limited by the emergence of real-time PCR in 1996.
  • dPCR has experienced a resurgence due to recent technological innovations.

Approach:

  • The technique involves partitioning a sample into numerous small reaction volumes.
  • Each partition is analyzed for the presence or absence of the target DNA.
  • Quantification is achieved by counting the positive partitions, enabling absolute quantification.

Key Points:

  • Digital PCR offers high precision and sensitivity for nucleic acid quantification.
  • Recent developments have simplified dPCR workflows and improved its practicality.
Keywords:
Digital PCRLimiting dilutionPCR

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  • This technique is valuable for applications such as rare mutation detection and copy number variation analysis.

    • Conclusions:

    • Digital PCR has undergone significant development and refinement.
    • Modern instrumentation and chemistry have overcome previous limitations, enhancing its usability.
    • The renewed accessibility of digital PCR positions it as a powerful tool in molecular diagnostics and research.