Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Portrayals of Food and Nutrition in Popular US Picture Books.

Pediatrics·2026

Same author

Maximum Matching Accuracy: An Instance Segmentation Evaluation Metric Utilizing Globally Optimal Matching.

ArXiv·2026

Same author

Membrane-resolved epithelial electrophysiology revealed using extracellular electrochemical impedance spectroscopy (EEIS).

bioRxiv : the preprint server for biology·2026

Same author

Mutation E300 recommended by protein language models gives ChrimsonR amplified photocurrent response.

bioRxiv : the preprint server for biology·2026

Same author

Regional alterations of parvalbumin interneurons in human App knock-in mouse model of Alzheimer's disease pathology.

Neurobiology of aging·2026

Same author

Sub-second extracellular impedance measurement of epithelial cell monolayers using step excitations and time-domain analysis.

Scientific reports·2026

Search research articles

Related Experiment Video

Updated: Apr 4, 2026

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Published on: May 22, 2012

Thermally multiplexed polymerase chain reaction.

Christopher R Phaneuf¹, Nikita Pak¹, D Curtis Saunders¹

¹George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology , Atlanta, Georgia 30332, USA.

Biomicrofluidics

|September 5, 2015

Summary

This summary is machine-generated.

Thermal multiplexing enables simultaneous amplification of multiple genetic targets in separate microfluidic reactions. This innovative polymerase chain reaction (PCR) method overcomes limitations of traditional techniques for efficient pathogen detection.

More Related Videos

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Published on: June 20, 2018

Rapid PCR Thermocycling using Microscale Thermal Convection

Rapid PCR Thermocycling using Microscale Thermal Convection

Published on: March 5, 2011

Related Experiment Videos

Last Updated: Apr 4, 2026

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Published on: May 22, 2012

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Published on: June 20, 2018

Rapid PCR Thermocycling using Microscale Thermal Convection

Rapid PCR Thermocycling using Microscale Thermal Convection

Published on: March 5, 2011

Area of Science:

Molecular Biology
Biotechnology
Microfluidics

Background:

Traditional polymerase chain reaction (PCR) methods for amplifying multiple genetic targets include serial reactions (time-consuming) and multiplex PCR (prone to bias and interactions).
There is a need for efficient, simultaneous amplification of diverse genetic targets under optimal conditions for each reaction.

Purpose of the Study:

To introduce and validate a novel thermocycling method called thermal multiplexing for simultaneous, independent amplification of multiple PCR targets.
To demonstrate the application of thermal multiplexing in a microfluidic system for pathogen detection.

Main Methods:

Developed a thermal multiplexing technique using a single, modulated heat source for independent temperature control of an array of PCR reactions.
Implemented the method using an infrared laser thermocycler and a microfluidic chip with 1 μl, oil-encapsulated reactions and closed-loop pulse-width modulation control.
Utilized heat transfer modeling to characterize system performance and validated it with simultaneous reactions at distinct annealing temperatures (48°C and 68°C).

Main Results:

Successfully demonstrated simultaneous amplification of two targets with widely different annealing temperatures, achieving excellent results.
Validated the thermal multiplexing method using clinical specimens, successfully amplifying and detecting both influenza A and B from human nasopharyngeal swabs.
Confirmed the scalability of thermal multiplexing for applications like broad-panel pathogen screening.

Conclusions:

Thermal multiplexing offers a scalable solution for simultaneous amplification of multiple genetic targets at their optimal conditions.
This method overcomes the limitations of serial and conventional multiplex PCR, improving efficiency and accuracy.
Thermal multiplexing shows significant potential for applications such as rapid and comprehensive pathogen detection in clinical settings.