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

BMC Biotechnology
L BogaertA Martens

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. 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. Based...Continue Reading

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Citations

Jun 23, 2012·The Journal of Cardiovascular Nursing·Ayman TailakhLorraine S Evangelista
Dec 17, 2009·BMC Molecular Biology·Yvonne KesslerRegina Hofmann-Lehmann
Feb 28, 2007·BMC Molecular Biology·Stefan ToegelHelmut Viernstein
Aug 19, 2007·BMC Molecular Biology·Ann-Britt NygardMerete Fredholm
Aug 20, 2008·BMC Molecular Biology·Aude Le BailBénédicte Charrier
Dec 4, 2013·BMC Veterinary Research·Lien BruynsteenMyriam Hesta
May 3, 2011·American Journal of Veterinary Research·Hugo HiltonJack R Snyder
Feb 1, 2013·American Journal of Veterinary Research·Kaitlyn A LinkBrandon N Lillie
Jan 15, 2014·Stem Cell Research & Therapy·Catharina De SchauwerGerlinde R Van de Walle
Dec 16, 2016·Journal of Equine Science·Sara Azarpeykan, Keren E Dittmer
Nov 1, 2008·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Siriporn PeansukmaneeAli Mobasheri
Aug 2, 2013·Reproduction, Fertility, and Development·Hilde NelisAnn Van Soom
Jul 13, 2020·Equine Veterinary Journal·Ewelina Semik-Gurgul
Sep 25, 2020·Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission·Özgecan Korkmaz AğaoğluSelim Aslan

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Methods Mentioned

BETA
reverse transcription PCR
PCR

Software Mentioned

Mfold
Primer
geNorm Visual Basic Excel
BLAST
geNorm

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