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Real Time RT-PCR

Real Time RT-PCR

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.
The real-time quantification of the number of amplified products is...

PCR

PCR

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Related Experiment Video

Updated: Sep 14, 2025

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format

Published on: August 20, 2018

Digital PCR: from early developments to its future application in clinics.

Amandine Trouchet¹, Guillaume Gines², Leonor Benhaim^1,3

¹Centre de Recherche des Cordeliers, UMRS1138, INSERM, CNRS, Sorbonne Université, USPC, Université Paris Cité, Equipe Labellisée Ligue Nationale Contre le Cancer, CNRS SNC 5096, Paris, France. valerie.taly@parisdescartes.fr.

|July 21, 2025

Summary

This summary is machine-generated.

Digital PCR (dPCR) offers precise, absolute quantification by partitioning samples into many reactions. This advanced technology provides high sensitivity and accuracy for diverse clinical applications, from cancer mutation detection to prenatal diagnostics.

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Area of Science:

Molecular Biology
Biotechnology
Clinical Diagnostics

Background:

Digital PCR (dPCR) represents the third generation of PCR technology.
It partitions samples into numerous parallel reactions for precise nucleic acid quantification.
This method relies on endpoint measurements and Poisson distribution for target concentration calculation.

Purpose of the Study:

To review the clinical applications of Digital PCR.
To highlight dPCR's advantages over existing technologies.
To provide an outlook on future developments in dPCR.

Main Methods:

Partitioning of PCR mixtures into a large number of parallel reactions.
Endpoint measurement of positive partitions.
Calculation of target concentration based on Poisson distribution.

Main Results:

dPCR offers high sensitivity, absolute quantification, accuracy, and reproducibility.
It enables detection of rare genetic mutations, crucial for oncology and liquid biopsies.
Applications extend to prenatal diagnosis and pathogen identification.

Conclusions:

Digital PCR is a powerful, calibration-free technology with broad clinical utility.
Its advantages are driving rapid adoption in research and diagnostics.
Future developments promise further expansion of dPCR's clinical impact.