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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: Jul 14, 2026

Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease
04:41

Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease

Published on: July 3, 2018

Quantitative (real-time) RT-PCR in cardiovascular research.

Kevin John Ashton1, John Patrick Headrick

  • 1Heart Foundation Research Centre, Griffith University, Southport, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|June 15, 2007
PubMed
Summary

Quantitative PCR (qPCR) offers sensitive, reproducible nucleic acid detection and quantification. This method is crucial for measuring mRNA levels in biological samples, especially in cardiovascular research.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cardiovascular Research

Background:

  • Quantitative PCR (qPCR) is a sensitive, specific, and reproducible method for nucleic acid detection.
  • Its large dynamic range and throughput make it ideal for quantifying mRNA levels.

Purpose of the Study:

  • To detail the methodology of qPCR experiments.
  • To provide guidance on applying qPCR in cardiovascular research.

Main Methods:

  • Sample preparation techniques.
  • Fluorescent chemistries for detection.
  • Primer and probe design principles.
  • Data analysis strategies for qPCR.

Main Results:

  • qPCR enables gel-free detection and quantification of nucleic acids.
  • The method reliably detects low template concentrations.
  • It offers a linear dynamic range up to 10 orders of magnitude.

Conclusions:

  • qPCR is a powerful tool for mRNA quantification in biological samples.
  • This chapter provides foundational knowledge for qPCR application in cardiovascular research.