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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...

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Related Experiment Video

Updated: Jun 3, 2026

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays
10:58

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Profiling microRNAs by real-time PCR.

Nana Jacobsen1, Ditte Andreasen, Peter Mouritzen

  • 1Exiqon A/S, Vedbaek, Denmark.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
PubMed
Summary
This summary is machine-generated.

Accurate microRNA (miRNA) quantification is crucial for research. This study details how primer strategies impact miRNA expression measurements and presents a locked nucleic acid real-time PCR method for reliable differential expression analysis.

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Related Experiment Videos

Last Updated: Jun 3, 2026

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays
10:58

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR
09:26

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

Published on: January 22, 2014

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
10:28

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNA (miRNA) expression changes are linked to physiological processes and diseases like cancer.
  • Accurate measurement of miRNA expression is a significant challenge in experimental systems.
  • Microarrays and real-time quantitative PCR (qPCR) are common methods for miRNA profiling.

Purpose of the Study:

  • To investigate the impact of different primer strategies on miRNA expression measurements.
  • To address the challenges of quantifying small miRNA molecules using PCR.
  • To demonstrate a reliable method for determining differentially expressed miRNAs.

Main Methods:

  • Evaluation of primer strategies for first-strand cDNA synthesis and PCR amplification.
  • Application of real-time quantitative PCR (qPCR).
  • Utilizing locked nucleic acid (LNA)-based qPCR for miRNA detection.

Main Results:

  • Primer design strategies significantly affect miRNA expression level measurements.
  • Standard PCR primer design guidelines are difficult to apply to short miRNA sequences.
  • The LNA-based real-time PCR approach effectively determines differentially expressed miRNAs.

Conclusions:

  • Optimizing primer strategies is essential for accurate miRNA expression analysis.
  • Novel approaches are needed to overcome the limitations of quantifying small RNA molecules.
  • Locked nucleic acid real-time PCR offers a robust method for differential miRNA expression studies.