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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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Related Experiment Video

Updated: Nov 8, 2025

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

A rapid and sensitive fluorescence biosensor based on plasmonic PCR.

Jingrui Wu¹, Kunlun Jiang, Hua Mi

¹Department of Chemistry, City University of Hong Kong, Hong Kong SAR, China. junghlee@cityu.edu.hk.

|April 23, 2021

Summary

This summary is machine-generated.

This study introduces plasmonic photothermal PCR (PPT-PCR), a rapid DNA detection method using magnetic nanoparticles. PPT-PCR achieves sensitive DNA target detection in just 5.5 minutes, overcoming limitations of traditional methods.

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Area of Science:

Biotechnology
Nanotechnology
Molecular Biology

Background:

Plasmonic PCR offers advantages in cost, size, and speed over conventional thermocyclers.
Conventional plasmonic PCR is hindered by fluorescence quenching, necessitating lengthy post-processing steps like centrifugation and gel electrophoresis, which negate rapid thermocycling benefits.

Purpose of the Study:

To develop a rapid and sensitive plasmonic photothermal PCR (PPT-PCR) assay with in situ end-point fluorescence detection.
To overcome the limitations of fluorescence quenching and post-processing steps in existing plasmonic PCR methods.

Main Methods:

Utilized plasmonic magnetic bi-functional nanoparticles for PCR amplification and detection.
Integrated photothermal effect with magnetic separation for streamlined sample processing.
Performed 30 thermocycles followed by fluorescence detection after magnetic separation.

Main Results:

Achieved DNA target detection within 5.5 minutes using the PPT-PCR assay.
Demonstrated a limit of detection comparable to real-time quantitative PCR (3.3 copies per μL).
Reduced overall assay time by approximately 5.5 times compared to conventional methods.

Conclusions:

The developed PPT-PCR assay provides a fast and sensitive method for DNA detection.
Combining photothermal effects and magnetic separation in a single nanoparticle system enhances PCR efficiency.
This strategy holds promise for developing advanced, rapid, and sensitive PCR-based biosensors for point-of-care testing.