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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.
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Related Experiment Video

Updated: Dec 14, 2025

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Published on: September 24, 2015

Fully automatic integrated continuous-flow digital PCR device for absolute DNA quantification.

Yangyang Jiang¹, Andreas Manz², Wenming Wu³

¹State Key Laboratory of Applied Optics, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, 130033, China.

Analytica Chimica Acta

|July 18, 2020

Summary

This summary is machine-generated.

A novel integrated digital polymerase chain reaction (dPCR) device automates the entire detection process in a single instrument. This innovation simplifies complex procedures and enhances efficiency for accurate hepatitis B virus quantification.

Keywords:

Contiuous flow Digital PCR Fully automatic Integrated device Microfluidics

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Last Updated: Dec 14, 2025

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Published on: September 24, 2015

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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Area of Science:

Biotechnology
Molecular Diagnostics
Bioengineering

Background:

Traditional digital polymerase chain reaction (dPCR) systems involve multiple separate instruments, leading to low integration, complex operations, and potential environmental contamination.
Existing dPCR workflows require extensive user intervention and multiple steps, hindering widespread adoption and increasing the risk of errors.

Purpose of the Study:

To develop and validate a fully automatic, integrated digital PCR device based on continuous-flow digital PCR theory.
To overcome the limitations of traditional dPCR instruments by creating a single, user-friendly system for automated molecular detection.

Main Methods:

Designed a fully automatic integrated digital PCR instrument incorporating sample injection, droplet generation, thermal cycling, and fluorescence detection.
Developed integrated control and data processing software for streamlined, one-step operation.
Utilized continuous-flow digital PCR principles for enhanced automation and efficiency.

Main Results:

The integrated device successfully automated all digital PCR procedures, from sample input to signal analysis.
Demonstrated the system's capability for absolute quantification of hepatitis B virus in serum samples across a dynamic range of 10^3 to 10^5 IU/mL.
Achieved a high linear correlation (0.9986) with traditional quantitative PCR (qPCR), confirming the instrument's reliability and accuracy.

Conclusions:

The proposed fully automatic integrated digital PCR device offers a significant advancement over traditional multi-instrument systems.
This innovative technology simplifies complex dPCR workflows, reduces operational difficulty, and minimizes environmental impact.
The system's solar-powered capability and high accuracy position it as a promising tool for promoting the development and application of integrated digital PCR systems.