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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...

PCR

PCR

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Related Experiment Video

Updated: Oct 28, 2025

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

Isothermal amplifications - a comprehensive review on current methods.

Jörn Glökler¹, Theam Soon Lim², Jeunice Ida²

¹Department of Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Wildau, Germany.

Critical Reviews in Biochemistry and Molecular Biology

|July 15, 2021

Summary

This summary is machine-generated.

Isothermal amplification methods offer rapid nucleic acid detection for medical diagnostics, advancing beyond traditional PCR. This review classifies and compares these techniques, aiding in selecting the best approach for various applications.

Keywords:

Isothermal amplification exponential DNA amplification methods nucleic acid-based diagnostics origin of amplification point-of-care diagnostics

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Rapid PCR Thermocycling using Microscale Thermal Convection

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Last Updated: Oct 28, 2025

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Rapid PCR Thermocycling using Microscale Thermal Convection

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Published on: March 5, 2011

Area of Science:

Molecular Biology
Medical Diagnostics
Biotechnology

Background:

Nucleic acid amplification techniques, particularly Polymerase Chain Reaction (PCR), have transformed molecular diagnostics.
Advancements in understanding DNA/RNA replication have led to the development of novel, temperature-independent amplification methods.
Isothermal amplification methods have emerged as a significant innovation in molecular diagnostics since the early 2000s.

Purpose of the Study:

To provide a comprehensive review of various isothermal nucleic acid amplification methods.
To classify these methods based on mechanistic characteristics, including reaction formats, amplification information, promoter, strand break, and refolding mechanisms.
To compare the efficiencies, usefulness, potential applications, and detection methods of different isothermal amplification strategies.

Main Methods:

Classification of isothermal amplification methods based on mechanistic properties.
Comparative analysis of the performance and utility of diverse isothermal amplification techniques.
Identification of potential applications and associated detection methodologies.

Main Results:

Isothermal amplification methods offer advantages in speed and simplicity for nucleic acid detection.
Heterogeneity in method-specific complications presents challenges in selecting optimal approaches.
A structured classification aids in understanding the landscape of isothermal amplification technologies.

Conclusions:

Isothermal amplification represents a significant advancement in molecular diagnostics, offering alternatives to temperature-cycling methods.
Understanding the diverse mechanisms and characteristics of isothermal methods is crucial for their effective application.
This review provides an outlook on the development and future potential of isothermal amplification techniques in diagnostics.