Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Tumor microenvironment-mediated interactions between macrophages and cancer cells define immunoregulatory transcriptional programs.

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie·2026

Same author

Personalized chemotherapy guidance using patient-derived scaffolds from peritoneal colorectal metastases.

Experimental hematology & oncology·2026

Same author

SiMSen-Seq STR sequencing across five laboratories enables lowered noise thresholds and improved allele calling.

Forensic science international. Genetics·2026

Same author

Understanding the qPCR Standard Curve: From Assay Validation to Absolute Quantification and Variance PCR.

International journal of molecular sciences·2026

Same author

Quantification Revisited: What qPCR Efficiency Models Reveal About Data Analysis Integrity.

International journal of molecular sciences·2026

Same author

Ageing promotes metastasis via activation of the integrated stress response.

Nature·2026

Search research articles

Related Experiment Video

Updated: May 2, 2026

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

The workflow of single-cell expression profiling using quantitative real-time PCR.

Anders Ståhlberg¹, Mikael Kubista

¹1Department of Pathology, Sahlgrenska Cancer Center, University of Gothenburg, Box 425, 40530 Gothenburg, Sweden.

Expert Review of Molecular Diagnostics

|March 22, 2014

Summary

This summary is machine-generated.

Single-cell profiling analyzes individual cells to reveal biological responses missed in bulk studies. Quantitative reverse transcription real-time PCR (RT-qPCR) is a powerful method for this analysis, with future improvements expected.

More Related Videos

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

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

Related Experiment Videos

Last Updated: May 2, 2026

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

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

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

Area of Science:

Biotechnology
Molecular Biology
Genomics

Background:

Biological samples are inherently heterogeneous, with diverse cellular responses to stimuli.
Classical bulk analysis often masks individual cell variations and responses.
Single-cell analysis offers a deeper understanding of biological complexity.

Purpose of the Study:

To review the current state of single-cell expression profiling techniques.
To highlight the advantages of single-cell analysis over traditional methods.
To discuss future advancements in single-cell profiling technologies.

Main Methods:

Quantitative reverse transcription real-time PCR (RT-qPCR) is the primary method for single-cell expression profiling.
Development of robust RT-qPCR workflows for high-throughput, sensitive, and specific measurements.
Application of RT-qPCR for analyzing mRNAs, microRNAs, non-coding RNAs, and proteins.

Main Results:

Single-cell profiling identifies cellular responses missed in bulk samples.
Enables discovery of novel cell types and subtypes.
Facilitates identification of key biological pathways and expression networks.

Conclusions:

Single-cell profiling, particularly using RT-qPCR, provides unprecedented biological insights.
Current RT-qPCR methods are highly sensitive, specific, and suitable for high-throughput analysis.
Significant improvements and developments in single-cell profiling are anticipated in the next five years.