Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

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

The WHO Classification of Genetic Tumour Syndromes: Considerations for Genetics.

Clinical genetics·2026
Same author

The WHO Classification of Genetic Tumour Syndromes: Considerations for histopathology.

Histopathology·2026
Same author

The International Classification of Diseases for Oncology, 4th Edition (ICD-O-4): An overview.

Cancer epidemiology·2026
Same author

Tumour markers and evidence-based pathology.

Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine·2026
Same author

A feasibility study using the ETL CoronaCheck® device to identify incident cases of SARS-CoV-2: FIND SARS-CoV-2.

Journal of breath research·2025
Same author

Development of a decision tree diagram for classifying study designs in tumour pathology research: a multidisciplinary approach.

The journal of pathology. Clinical research·2025

Related Experiment Video

Updated: Jun 2, 2026

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
10:44

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

Published on: June 20, 2018

Measuring gene expression from cell cultures by quantitative reverse-transcriptase polymerase chain reaction.

Sharon Glaysher1, Francis G Gabriel, Ian A Cree

  • 1Translational Oncology Research Centre, Queen Alexandra Hospital, Portsmouth, UK.

Methods in Molecular Biology (Clifton, N.J.)
|April 26, 2011
PubMed
Summary

Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a reliable method for measuring gene expression in cells. This guide details RNA extraction and the two-step qRT-PCR process for accurate gene analysis.

More Related Videos

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
08:36

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction

Published on: November 1, 2018

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
10:50

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

Published on: February 25, 2017

Related Experiment Videos

Last Updated: Jun 2, 2026

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
10:44

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

Published on: June 20, 2018

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
08:36

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction

Published on: November 1, 2018

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
10:50

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

Published on: February 25, 2017

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a fundamental technique in molecular biology.
  • Accurate measurement of RNA levels is crucial for understanding gene expression and cellular function.

Purpose of the Study:

  • To provide a comprehensive overview of the quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) method.
  • To outline the essential steps involved in RNA extraction and the two-step qRT-PCR process.
  • To guide researchers in the analysis of qRT-PCR results.

Main Methods:

  • RNA extraction from cellular samples.
  • Two-step qRT-PCR, involving reverse transcription to synthesize complementary DNA (cDNA) followed by polymerase chain reaction (PCR) amplification.
  • Analysis of PCR product quantification.

Main Results:

  • Successful RNA extraction and cDNA synthesis are critical for reliable qRT-PCR outcomes.
  • Commercial kits can facilitate RNA extraction and cDNA synthesis.
  • Ensuring adequate cDNA quality and quantity is essential for downstream applications like standard PCR or arrays.

Conclusions:

  • qRT-PCR is a powerful and versatile tool for quantifying gene expression.
  • Proper execution of RNA extraction and the two-step qRT-PCR protocol is vital for accurate results.
  • Attention to cDNA quality and quantity is paramount for successful gene expression analysis.