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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 30, 2026

Absorbent Microbiopsy Sampling and RNA Extraction for Minimally Invasive, Simultaneous Blood and Skin Analysis
06:18

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Reference genes for canine skin when using quantitative real-time PCR.

Shona H Wood1, Dylan N Clements, Neil A McEwan

  • 1Department of Veterinary Pathology, Faculty of Veterinary Science, University of Liverpool, Liverpool, UK. shona.wood@liv.ac.uk

Veterinary Immunology and Immunopathology
|October 1, 2008
PubMed
Summary
This summary is machine-generated.

Quantitative real-time PCR (qPCR) requires stable reference genes for accurate mRNA quantification. This study found RPL13A and CG14980 are stable in canine skin, unlike the commonly used GAPDH.

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Area of Science:

  • Veterinary Science
  • Molecular Biology
  • Genomics

Background:

  • Quantitative real-time PCR (qPCR) is crucial for mRNA expression analysis.
  • Accurate qPCR relies on normalizing data with stable reference genes.
  • GAPDH is widely used but often shows unstable expression across tissues.

Purpose of the Study:

  • To evaluate the stability of seven potential reference genes in canine skin.
  • To identify suitable reference genes for qPCR studies in canine skin, including atopic dermatitis models.

Main Methods:

  • Seven reference genes (IMP, CG14980, S7, HIRA, GAPDH, RPL13A, SDHA) were quantified in canine skin samples (healthy, lesional, non-lesional atopic dermatitis).
  • Reference gene stability was assessed using three statistical algorithms: Bestkeeper, GeNorm, and Normfinder.

Main Results:

  • The study confirmed that GAPDH is not a stably expressed reference gene in canine skin.
  • RPL13A and CG14980 demonstrated the highest expression stability in canine whole skin.
  • This finding impacts the interpretation of previous qPCR studies using GAPDH in canine skin.

Conclusions:

  • GAPDH is unsuitable as a reference gene for qPCR in canine skin.
  • RPL13A and CG14980 are recommended as reliable reference genes for future qPCR studies in canine skin.
  • This research provides essential guidance for accurate gene expression analysis in canine dermatology.