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Real Time RT-PCR

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

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Related Experiment Video

Updated: Jul 17, 2025

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

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Published on: September 24, 2015

Label-free DNA quantification using isothermal amplification on an exposed core optical fiber microfluidic platform.

Xuegang Li^1,2, He Zhang¹, Yanan Zhang^1,2

¹College of Information Science and Engineering, Northeastern University, Shenyang, Liaoning 110819, China. zhouxue@ise.neu.edu.cn.

|September 1, 2023

Summary

This study introduces a novel, label-free method for detecting DNA using optical fiber sensing and isothermal amplification. The technology offers rapid, sensitive, and portable deoxyribonucleic acid (DNA) quantification, crucial for disease outbreak management.

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Area of Science:

Biomedical Engineering
Analytical Chemistry
Optoelectronics

Background:

Isothermal amplification offers efficient DNA detection compatible with portable devices.
The COVID-19 pandemic highlighted the need for rapid, widespread diagnostic tools.
Existing methods may lack the sensitivity, speed, or portability required for field applications.

Purpose of the Study:

To develop and demonstrate a novel, label-free isothermal deoxyribonucleic acid (DNA) amplification method.
To integrate optical fiber sensing with microfluidics for real-time DNA quantification.
To achieve highly sensitive and selective DNA detection using refractive index (RI) changes.

Main Methods:

Fabrication of a Mach-Zehnder (MZ) interference structure using exposed-core fiber (ECF).
Integration of a microfluidic channel for sample and amplification solution delivery.
Real-time monitoring of solution RI changes during isothermal DNA amplification.

Main Results:

Successful demonstration of a label-free, real-time quantitative measurement of DNA amplification.
Detection of ultra-low DNA concentrations (0.16 aM) using the proposed platform.
Validation of the system's high sensitivity, selectivity, and portability.

Conclusions:

The developed optical fiber-based platform provides a breakthrough for label-free DNA detection and quantification.
This method meets the demand for simple, rapid, portable, and highly sensitive diagnostic tools.
Potential applications include point-of-care diagnostics and environmental monitoring.