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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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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: Nov 10, 2025

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Stem-Loop qRT-PCR-Based Quantification of miRNAs.

Yoann Abel1, Mathieu Rederstorff2

  • 1IGMM, CNRS, Université de Montpellier, Montpellier, France. yoann.abel@igmm.cnrs.fr.

Methods in Molecular Biology (Clifton, N.J.)
|April 1, 2021
PubMed
Summary
This summary is machine-generated.

Stem-loop quantitative reverse transcription PCR (qRT-PCR) is a common method for quantifying microRNAs (miRNAs). This technique uses a two-step process involving reverse transcription and real-time PCR for sensitive and specific miRNA analysis.

Keywords:
MicroRNAReal-time PCR quantificationSmall non-coding RNAStem-loop primers

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Small non-coding RNAs, particularly microRNAs (miRNAs), play crucial roles in gene regulation.
  • Accurate quantification of miRNAs is essential for understanding their biological functions and involvement in diseases.
  • Quantitative reverse transcription PCR (qRT-PCR) is a widely adopted method for RNA analysis.

Purpose of the Study:

  • To describe the stem-loop qRT-PCR method for quantifying small non-coding RNAs like miRNAs.
  • To highlight the advantages of this method in terms of speed, simplicity, sensitivity, and specificity.

Main Methods:

  • The stem-loop qRT-PCR method involves a two-step quantitative analysis.
  • Step 1: RNA is reverse transcribed using a specific stem-loop primer.
  • Step 2: The resulting complementary DNA (cDNA) is used as a template for quantitative real-time PCR.

Main Results:

  • The stem-loop qRT-PCR method enables sensitive and specific quantification of miRNAs.
  • This approach provides reliable quantitative data for miRNA expression analysis.
  • The method is characterized by its speed and simplicity.

Conclusions:

  • Stem-loop qRT-PCR is a highly effective and commonly used approach for quantifying miRNAs.
  • This technique facilitates the study of miRNA functions due to its high sensitivity and specificity.
  • The method offers a fast and straightforward way to obtain quantitative miRNA data.