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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...

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Updated: Mar 17, 2026

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

Published on: March 9, 2016

The Next-Generation PCR-Based Quantification Method for Ambient Waters: Digital PCR.

Yiping Cao¹, John F Griffith¹, Stephen B Weisberg²

¹Southern California Coastal Water Research Project Authority, 3535 Harbor Blvd, Costa Mesa, CA, 92626, USA.

Methods in Molecular Biology (Clifton, N.J.)

|July 28, 2016

Summary

This summary is machine-generated.

Digital polymerase chain reaction (digital PCR) offers advanced DNA quantification for water monitoring. This method refines real-time quantitative PCR (qPCR) by eliminating standard curves and improving accuracy for field applications.

Keywords:

Ambient water monitoring Beach water quality Droplet digital PCR Multiplex Polymerase chain reaction

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Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

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Published on: June 11, 2016

Area of Science:

Environmental science
Molecular biology
Analytical chemistry

Background:

Real-time quantitative PCR (qPCR) is a common method for ambient water monitoring.
Digital polymerase chain reaction (digital PCR) represents an advancement over qPCR.

Purpose of the Study:

To highlight the advantages of digital PCR for ambient water monitoring.
To explain the principles and benefits of digital PCR compared to qPCR.

Main Methods:

Digital PCR partitions samples into numerous miniature reactions for individual analysis.
DNA density is calculated using Poisson statistics based on the fraction of positive reactions.
This method bypasses the need for standard curves, unlike traditional qPCR.

Main Results:

Digital PCR eliminates the labor and variability associated with standard curves.
It offers improved performance, including reduced inhibition susceptibility, enhanced repeatability, and reproducibility.
The technique allows for multiplexing, enabling the measurement of multiple targets simultaneously.

Conclusions:

Digital PCR is highly suitable for ambient water monitoring applications.
Its advantages make it particularly valuable for autonomous field-based molecular monitoring.