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Home / Equine Research Bank / BMC Biotechnol / Selection of a set of reliable reference genes for quantitat...
BMC biotechnology2006; 6; 24; doi: 10.1186/1472-6750-6-24

Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids.

Abstract: Real-time quantitative PCR can be a very powerful and accurate technique to examine gene transcription patterns in different biological conditions. One of the critical steps in comparing transcription profiles is accurate normalisation. In most of the studies published on real-time PCR in horses, normalisation occurred against only one reference gene, usually GAPDH or ACTB, without validation of its expression stability. This might result in unreliable conclusions, because it has been demonstrated that the expression levels of so called "housekeeping genes" may vary considerably in different tissues, cell types or disease stages, particularly in clinical samples associated with malignant disease. The goal of this study was to establish a reliable set of reference genes for studies concerning normal equine skin and equine sarcoids, which are the most common skin tumour in horses. Results: In the present study the gene transcription levels of 6 commonly used reference genes (ACTB, B2M, HPRT1, UBB, TUBA1 and RPL32) were determined in normal equine skin and in equine sarcoids. After applying the geNorm applet to this set of genes, TUBA1, ACTB and UBB were found to be most stable in normal skin and B2M, ACTB and UBB in equine sarcoids. Conclusions: Based on these results, TUBA1, ACTB and UBB, respectively B2M, ACTB and UBB can be proposed as reference gene panels for accurate normalisation of quantitative data for normal equine skin, respectively equine sarcoids. When normal skin and equine sarcoids are compared, the use of the geometric mean of UBB, ACTB and B2M can be recommended as a reliable and accurate normalisation factor.
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Publication Date: 2006-04-27 PubMed ID: 16643647PubMed Central: PMC1484482DOI: 10.1186/1472-6750-6-24Google Scholar: Lookup
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  • Journal Article
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article focuses on identifying a reliable set of reference genes to accurately measure gene transcription levels in normal equine skin and in equine sarcoids, a common skin tumor in horses, using real-time quantitative PCR technique.

Objective and Importance of the Study

  • The primary aim of the research was to pinpoint dependable reference genes for real-time quantitative PCR studies relating to normal equine skin and equine sarcoids.
  • Equine sarcoids, which are the most prevalent skin tumors in horses, were of particular interest.
  • A crucial aspect of comparing transcript profiles is accurate normalization, often done against a single reference gene like GAPDH or ACTB, without validating its expression stability. This approach could lead to unreliable results as “housekeeping genes” expression can considerablely vary in different tissues, cell types or disease stages.

Investigation and Results Obtained

  • Six prevalent reference genes (ACTB, B2M, HPRT1, UBB, TUBA1, and RPL32) were the focus of the investigation. The researchers measured gene transcription levels in normal equine skin and in equine sarcoids using these genes.
  • The geNorm applet was employed to these genes to assess their stability. TUBA1, ACTB, and UBB were discovered to be the most stable in normal skin, while B2M, ACTB, and UBB were found the most stable in equine sarcoids.

Conclusions and Recommendations

  • Based on the study findings, TUBA1, ACTB, and UBB for normal equine skin, and B2M, ACTB, and UBB for equine sarcoids could be recommended as reference gene panels for accurate normalization of quantitative data.
  • When comparing normal skin and equine sarcoids, the geometric mean of UBB, ACTB, and B2M is recommended as a reliable and accurate normalization factor.

Cite This Article

APA
Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A. (2006). Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids. BMC Biotechnol, 6, 24. https://doi.org/10.1186/1472-6750-6-24

Publication

BMC biotechnology
ISSN: 1472-6750
NlmUniqueID: 101088663
Country: England
Language: English
Volume: 6
Pages: 24

Researcher Affiliations

Bogaert, Lies
  • Department of Surgery and Anaesthesiology of Domestic Animals, Faculty of Veterinary Medicine, Ghent University--UGent, Salisburylaan 133, B-9820 Merelbeke, Belgium. lies.bogaert@UGent.be
Van Poucke, Mario
    De Baere, Cindy
      Peelman, Luc
        Gasthuys, Frank
          Martens, Ann

            MeSH Terms

            • Animals
            • Gene Expression Profiling
            • Genes
            • Horse Diseases / genetics
            • Horse Diseases / pathology
            • Horses
            • Polymerase Chain Reaction / methods
            • Polymerase Chain Reaction / veterinary
            • Skin / pathology
            • Skin Neoplasms / genetics
            • Skin Neoplasms / pathology
            • Skin Neoplasms / veterinary
            • Transcription, Genetic

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