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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.
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Related Experiment Video

Updated: Apr 16, 2026

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Stem-loop RT-PCR based quantification of small non-coding RNAs.

Véronique Salone1, Mathieu Rederstorff

  • 1CNRS UMR 7365, IMoPA, Université de Lorraine, 9 avenue de la Forêt de Haye, CS 50184, Vandoeuvre-lès-Nancy, 54506, France.

Methods in Molecular Biology (Clifton, N.J.)
|March 21, 2015
PubMed
Summary
This summary is machine-generated.

Stem-loop quantitative reverse transcription PCR (qRT-PCR) is a common method for quantifying microRNAs (miRNAs). This sensitive and specific technique involves reverse transcription followed by real-time PCR for accurate small RNA analysis.

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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.
  • Traditional RNA quantification methods may lack the sensitivity and specificity required for small RNAs.

Purpose of the Study:

  • To describe and highlight the utility of the stem-loop quantitative reverse transcription PCR (qRT-PCR) method.
  • To emphasize its application in the sensitive and specific quantification of microRNAs.
  • To showcase its role in studying miRNA functions.

Main Methods:

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

Main Results:

  • The stem-loop qRT-PCR approach provides highly sensitive quantification of small non-coding RNAs.
  • The method demonstrates high specificity, crucial for distinguishing between similar miRNA sequences.
  • It offers a fast and straightforward protocol for miRNA analysis.

Conclusions:

  • Stem-loop qRT-PCR is an effective and widely adopted technique for miRNA quantification.
  • Its sensitivity and specificity make it ideal for studying miRNA expression and function.
  • This method facilitates research into the roles of miRNAs in various biological processes.