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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...

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Related Experiment Video

Updated: May 27, 2026

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR
10:23

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Published on: February 12, 2018

Quantification of transcript levels with quantitative RT-PCR.

Karen L Carleton1

  • 1University of Maryland, College Park, MD, USA. kcarleto@umd.edu

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2011
PubMed
Summary
This summary is machine-generated.

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) confirms gene expression differences between organisms. This method involves RNA extraction, cDNA synthesis, and gene expression quantification for accurate phenotypic divergence studies.

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Related Experiment Videos

Last Updated: May 27, 2026

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR
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Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR
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Area of Science:

  • Molecular Biology
  • Genetics
  • Evolutionary Biology

Background:

  • Differential gene expression drives phenotypic divergence between species.
  • Accurate quantification of gene expression is crucial for understanding evolutionary processes.
  • High-throughput methods can identify candidate genes, but definitive confirmation is needed.

Purpose of the Study:

  • To describe the definitive method for confirming gene expression differences.
  • To outline the essential steps for performing quantitative reverse transcription polymerase chain reaction (qRT-PCR).

Main Methods:

  • Extraction of total RNA from biological samples.
  • Reverse transcription of RNA to complementary DNA (cDNA).
  • Quantification of relative gene expression using real-time quantitative PCR (qPCR).

Main Results:

  • The described protocol provides a quantitative measure of gene expression.
  • This method allows for the confirmation of candidate gene expression differences identified by large-scale approaches.

Conclusions:

  • Quantitative RT-PCR is the definitive technique for validating gene expression divergence.
  • This methodology is essential for studying the genetic basis of phenotypic differences.