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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 22, 2026

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate
11:13

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

Published on: May 9, 2019

A novel and universal method for microRNA RT-qPCR data normalization.

Pieter Mestdagh1, Pieter Van Vlierberghe, An De Weer

  • 1Center for Medical Genetics, Ghent University Hospital, De Pintelaan 185, Ghent, Belgium. pieter.mestdagh@ugent.be

Genome Biology
|June 18, 2009
PubMed
Summary

Analyzing microRNA expression is crucial. This study shows using the mean microRNA expression value normalizes data better than current methods, reducing technical noise and improving biological insights for microRNA real-time quantitative PCR.

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Published on: August 3, 2011

Related Experiment Videos

Last Updated: Jun 22, 2026

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate
11:13

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

Published on: May 9, 2019

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

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
07:27

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNA (miRNA) gene expression analysis is vital in biological research.
  • Accurate normalization is essential for reliable miRNA real-time quantitative PCR (qPCR) data.
  • Current normalization strategies may not fully account for technical variations.

Purpose of the Study:

  • To evaluate the mean expression value of all expressed miRNAs as a normalization factor for miRNA qPCR.
  • To compare this novel approach against the currently adopted normalization strategy.
  • To assess the impact on reducing technical variation and improving biological change detection.

Main Methods:

  • MicroRNA gene expression profiling using real-time quantitative PCR.
  • Calculation of the mean expression value across all detected miRNAs per sample.
  • Comparative analysis of data normalized by the mean expression value versus a standard method.

Main Results:

  • The mean expression value normalization significantly reduced technical variation in miRNA qPCR data.
  • This method provided a more accurate assessment of genuine biological differences between samples.
  • Performance was superior to the currently adopted normalization approach.

Conclusions:

  • The mean expression value of all expressed miRNAs is a robust and effective normalization factor for miRNA qPCR.
  • This strategy enhances data reliability by minimizing technical noise.
  • It facilitates more accurate identification and interpretation of biological variations in miRNA expression studies.