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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: Mar 30, 2026

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants

Published on: August 5, 2020

Quantitative Reverse Transcription-qPCR-Based Gene Expression Analysis in Plants.

Heithem Ben Abdallah¹, Petra Bauer^2,3

¹Institute of Botany, Heinrich-Heine University, Universitätstrasse 1, 40225, Düsseldorf, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|November 19, 2015

Summary

This study provides a reliable protocol for reverse transcription-quantitative real-time PCR (RT-qPCR) assays in plants. This method enables accurate gene expression analysis in any plant species, even without genomic data.

Keywords:

Cq value Gene expression Primer design Reference gene Reverse transcription cDNA qPCR

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Published on: January 11, 2017

Area of Science:

Plant molecular biology
Gene expression analysis
Biotechnology

Background:

Understanding gene function requires studying gene expression patterns.
Reverse transcription-quantitative real-time PCR (RT-qPCR) is a fast, reproducible, and quantitative method for gene expression analysis.
RT-qPCR can be applied to diverse plant species, including non-model organisms, without prior genomic information.

Purpose of the Study:

To provide a detailed and adaptable protocol for performing reliable RT-qPCR assays in plants.
To facilitate gene expression studies across various plant species.

Main Methods:

Detailed protocol development for RT-qPCR.
Optimization of qPCR strategy and primer design.
Guidelines for plant material preparation, RNA extraction, and cDNA synthesis.
Standardized procedures for qPCR setup, execution, and data analysis.

Main Results:

A comprehensive and adaptable protocol for plant RT-qPCR is presented.
The protocol covers all essential steps from experimental design to data interpretation.
The method is suitable for both model and non-model plant species.

Conclusions:

This protocol offers a robust framework for accurate gene expression quantification in plants.
The presented method is highly adaptable, supporting research in diverse plant systems.
It empowers researchers to investigate gene function effectively, even with limited genomic resources.