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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: Aug 30, 2025

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Published on: June 14, 2013

Label and quantify mRNA molecules in live cell experiments using SunRISER and dNEMO.

Yue Guo¹, Gabriel J Kowalczyk², Robin E C Lee³

¹Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15260, USA; Department of Physics and Astronomy, University of Pittsburgh, Pittsburgh, PA 15260, USA.

|August 29, 2022

Summary

This study presents a new protocol for visualizing messenger RNA (mRNA) in single cells over extended periods. The method enables detailed analysis of mRNA dynamics, offering insights into cellular heterogeneity.

Keywords:

Bioinformatics Biotechnology and bioengineering Microscopy Molecular biology Molecular/Chemical probes

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Last Updated: Aug 30, 2025

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Published on: June 14, 2013

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

Published on: June 4, 2019

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Published on: December 21, 2017

Area of Science:

Molecular Biology
Cell Biology
Biotechnology

Background:

Visualization of mRNA in single cells is crucial for understanding gene expression.
Existing methods face limitations in temporal resolution and duration.
Cell-to-cell and spatiotemporal heterogeneity in mRNA distribution are key areas of study.

Purpose of the Study:

To describe a novel protocol for labeling, visualizing, and quantifying mRNA molecules.
To enable time-lapse imaging of mRNA over hours to days.
To provide resources for implementing the protocol.

Main Methods:

Development of a protocol for mRNA labeling in single cells.
Application of time-lapse imaging for extended observation.
Utilizing free software for semi-automated image analysis.

Main Results:

The protocol allows for the visualization and quantification of mRNA molecules.
It enables observation of mRNA dynamics over durations of hours to days.
The method provides a semi-automated pipeline for image analysis.

Conclusions:

The described protocol offers a powerful tool for studying mRNA dynamics in single cells.
It facilitates the investigation of cellular heterogeneity at the molecular level.
The protocol is supported by readily available plasmids and analysis software.