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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: Oct 15, 2025

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Published on: January 19, 2017

Tutorial: Guidelines for Single-Cell RT-qPCR.

Daniel Zucha^1,2, Mikael Kubista^1,3, Lukas Valihrach¹

¹Laboratory of Gene Expression, Institute of Biotechnology CAS, 252 50 Vestec, Czech Republic.

|October 23, 2021

Summary

This summary is machine-generated.

Single-cell reverse transcription quantitative PCR (scRT-qPCR) is a precise method for gene expression analysis. This tutorial guides researchers through optimizing scRT-qPCR for reliable single-cell gene expression studies.

Keywords:

RT-qPCR gene expression preamplification quantitative PCR reverse transcription sample collection single cell

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Last Updated: Oct 15, 2025

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Published on: January 19, 2017

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

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Published on: June 20, 2018

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Reverse transcription quantitative PCR (RT-qPCR) is crucial for gene expression analysis.
RT-qPCR offers precision, sensitivity, flexibility, and cost-effectiveness.
Single-cell RT-qPCR (scRT-qPCR) is valuable for screening and validation in gene expression studies.

Purpose of the Study:

To provide a comprehensive summary of the scRT-qPCR method.
To offer guidelines for performing high-standard scRT-qPCR measurements.
To address limitations and provide recommendations for optimizing scRT-qPCR.

Main Methods:

Discussion of single-cell collection method limitations.
Emphasis on the importance of the reverse transcription step.
Recommendations for preamplification and primer design.
Summary of essential data processing steps.

Main Results:

scRT-qPCR is a well-established method for gene expression analysis.
The method is suitable for pre-screening before scRNA-Seq or validating high-throughput findings.
Detailed protocol guidelines are provided for researchers.

Conclusions:

This tutorial offers a comprehensive guide to scRT-qPCR.
It empowers researchers to perform high-standard single-cell gene expression measurements.
The guidelines cover critical steps from cell collection to data processing.