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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: May 15, 2026

Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis
07:29

Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis

Published on: May 16, 2020

MicroRNA profiling during human keratinocyte differentiation using a quantitative real-time PCR method.

Janosch Hildebrand1, Adam Grundhoff, Stefan Gallinat

  • 1R&D, Skin Research Center, Beiersdorf AG, Hamburg, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|January 18, 2013
PubMed
Summary

MicroRNAs (miRNAs) regulate epidermal keratinocyte differentiation. A new stem loop real-time PCR method enables reliable miRNA profiling in limited human skin samples, including epidermal stem cells.

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Last Updated: May 15, 2026

Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis
07:29

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10:48

Differentiation of a Human Neural Stem Cell Line on Three Dimensional Cultures, Analysis of MicroRNA and Putative Target Genes

Published on: April 12, 2015

Area of Science:

  • Dermatology
  • Molecular Biology
  • Genetics

Background:

  • Epidermal keratinocyte terminal differentiation involves complex transcriptional and posttranscriptional regulation.
  • MicroRNAs (miRNAs) are key posttranscriptional regulators impacting cellular differentiation processes.
  • Existing miRNA detection methods like northern blotting, microarrays, and deep sequencing have limitations in sensitivity, cost, or applicability to scarce samples.

Purpose of the Study:

  • To describe a general real-time PCR method for miRNA profiling.
  • To demonstrate the method's utility for analyzing miRNA expression in various human skin cell types, including limited samples.

Main Methods:

  • Stem loop primer-based real-time PCR for simultaneous miRNA detection.
  • Application of the method to isolated epidermal stem cells, transiently amplifying cells, and terminally differentiated keratinocytes from human skin.

Main Results:

  • The described real-time PCR method allows for fast, specific, and reliable miRNA profiling.
  • This technique is effective even for samples with limited availability, such as human epidermal stem cells.

Conclusions:

  • Stem loop real-time PCR offers a robust and accessible approach for miRNA profiling in human skin.
  • This method facilitates the study of miRNA roles in epidermal differentiation and diseases, particularly with limited clinical samples.