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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: Apr 20, 2026

Author Spotlight: Advancing Rapid Detection of Respiratory Pathogens Using Microfluidic Chip

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Published on: March 29, 2024

Microfluidic continuous flow digital loop-mediated isothermal amplification (LAMP).

Tushar D Rane¹, Liben Chen, Helena C Zec

¹Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA. thwang@jhu.edu.

|November 29, 2014

Summary

This summary is machine-generated.

This study introduces a novel microfluidic droplet device for streamlined digital nucleic acid detection using loop-mediated isothermal amplification (LAMP). This integrated system simplifies workflows and enhances throughput for ultra-sensitive molecular diagnostics.

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Area of Science:

Biomedical engineering
Molecular diagnostics
Microfluidics

Background:

Digital nucleic acid detection is crucial for sensitive molecular analysis in biomedical research.
Digital polymerase chain reaction (dPCR) is widely used but requires complex thermocycling, hindering microfluidic integration.
Loop-mediated isothermal amplification (LAMP) offers a promising isothermal alternative with high specificity.

Purpose of the Study:

To develop an integrated microfluidic droplet device for digital nucleic acid detection.
To implement all necessary steps—droplet generation, incubation, and detection—on a single platform.
To overcome the limitations of existing digital PCR and microchamber-based assays.

Main Methods:

A microfluidic droplet device was engineered for continuous flow operation.
The device integrates droplet generation, isothermal amplification (LAMP), and in-line detection.
The system performs digital LAMP assays within discrete microfluidic droplets.

Main Results:

The device successfully performs digital nucleic acid detection using LAMP in a single, continuous flow system.
Continuous flow operation removes limitations on reaction numbers and improves analysis throughput compared to static digital assays.
The integrated approach simplifies complex workflows associated with traditional digital PCR.

Conclusions:

The developed microfluidic droplet device offers a streamlined and efficient platform for digital LAMP assays.
This technology has the potential to advance ultra-sensitive and quantitative nucleic acid detection in various biomedical applications.
The integrated, continuous flow design represents a significant improvement over fragmented, multi-instrument workflows.