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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: May 9, 2026

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

Published on: February 3, 2021

Simple system for isothermal DNA amplification coupled to lateral flow detection.

Kristina Roskos¹, Anna I Hickerson, Hsiang-Wei Lu

¹Keck Graduate Institute of Applied Life Sciences, Claremont, California, United States of America.

|August 8, 2013

Summary

This study presents a low-cost, automated nucleic acid testing system for infectious diseases. The portable device uses a mesofluidic cartridge for rapid DNA amplification and detection, improving point-of-care diagnostics.

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Last Updated: May 9, 2026

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

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Rapid Detection of Bacterial Pathogens Causing Lower Respiratory Tract Infections via Microfluidic-Chip-Based Loop-Mediated Isothermal Amplification

Published on: March 29, 2024

Area of Science:

Biomedical Engineering
Molecular Diagnostics
Point-of-Care Testing

Background:

Integrated, automated nucleic acid testing is crucial for improving infectious disease diagnosis in point-of-care settings.
Current methods often lack the integration and automation needed for rapid, accessible diagnostics.

Purpose of the Study:

To develop an early prototype of a low-cost, automated system for infectious disease diagnosis.
To demonstrate the feasibility of isothermal DNA amplification coupled with nucleic acid lateral flow (NALF) detection in a mesofluidic cartridge.

Main Methods:

A mesofluidic cartridge system utilizing passive valves, flexible pouches, and electrolysis-driven pumps for fluid handling.
Isothermal DNA amplification (Loop Mediated Amplification or Exponential Amplification Reaction) coupled to NALF detection.
Demonstration of Mycobacterium tuberculosis (M.tb) genomic DNA amplification and detection.

Main Results:

Successful amplification and detection of Mycobacterium tuberculosis genomic DNA using the developed system.
The cartridge design facilitates compact, inexpensive instrument development and prevents amplicon contamination.
Demonstrated proof-of-principle for isothermal amplification and NALF detection within the mesofluidic cartridge.

Conclusions:

The developed low-cost system shows promise for automated, sample-in to answer-out infectious disease diagnosis.
Refined versions could enable fully-automated diagnostics in primary care settings, particularly in low-resource countries.
The scalable cartridge design supports potential for widespread adoption in resource-limited areas.