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Int. J. Mol. Sci. 2011, 12(3), 1727-1734; doi:10.3390/ijms12031727

Evaluation of Potential Reference Genes for Relative Quantification by RT-qPCR in Different Porcine Tissues Derived from Feeding Studies

1
Institute of Animal Nutrition, Products, and Nutrition Physiology (APN), University of Natural Resources and Life Sciences, 1190 Vienna, Austria
2
Department of Food Sciences and Technology, Institute for Food Science, University of Natural Resources and Life Sciences, 1190 Vienna, Austria
3
Chair of Animal Nutrition, Technische Universitaet Muenchen-Weihenstephan, Liesel-Beckmann-Strasse 6, 85350 Freising-Weihenstephan, Germany
*
Author to whom correspondence should be addressed.
Received: 24 December 2010 / Revised: 29 January 2011 / Accepted: 25 February 2011 / Published: 7 March 2011
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract

Five potential reference genes for RT-qPCR application, namely histone H3, beta-actin, GAPDH, ubiquitin and 18S rRNA, were evaluated for normalization of gene expression in four selected tissues (liver, kidney, thyroid and abdominal fat). Tissues were derived from fattening pigs exposed to different amounts and type of dietary iodine. Two software applications (geNorm and NormFinder) were used to evaluate the stability of the potential reference genes. All studied genes displayed high expression stability but different stability patterns between the investigated tissues. The results suggest GAPDH and 18S rRNA as reference genes applicable in all tissues investigated. Beta-actin and histone H3 are suitable reference genes for all tissues investigated except fat. In contrast, ubiquitin should be excluded from use as a reference gene in the porcine tissues analyzed due to variations in expression levels, despite the good expression stability.
Keywords: sus scrofa; RT-qPCR; reference genes; gene stability sus scrofa; RT-qPCR; reference genes; gene stability
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Li, Q.; Domig, K.J.; Ettle, T.; Windisch, W.; Mair, C.; Schedle, K. Evaluation of Potential Reference Genes for Relative Quantification by RT-qPCR in Different Porcine Tissues Derived from Feeding Studies. Int. J. Mol. Sci. 2011, 12, 1727-1734.

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