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Selection of Internal Reference Genes for Normalization of Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) Analysis in the Canine Brain and Other Organs

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Abstract

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a specific and sensitive technique for quantifying gene expression. To analyze qRT-PCR data accurately, suitable reference genes that show consistent expression patterns across different tissues and experimental conditions should be selected. The objective of this study was to obtain the most stable reference genes in dogs, using samples from 13 different brain tissues and 10 other organs. 16 well-known candidate reference genes were analyzed by the geNorm, NormFinder, and BestKeeper programs. Brain tissues were derived from several different anatomical regions, including the forebrain, cerebrum, diencephalon, hindbrain, and metencephalon, and grouped accordingly. Combination of the three different analyses clearly indicated that the ideal reference genes are ribosomal protien S5 (RPS5) in whole brain, RPL8 and RPS5 in whole body tissues, RPS5 and RPS19 in the forebrain and cerebrum, RPL32 and RPS19 in the diencephalon, GAPDH and RPS19 in the hindbrain, and MRPS7 and RPL13A in the metencephalon. These genes were identified as ideal for the normalization of qRT-PCR results in the respective tissues. These findings indicate more suitable and stable reference genes for future studies of canine gene expression.

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Acknowledgments

This research was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea (AGC1021112).

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Authors and Affiliations

  1. National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Chungbuk, 363-883, Republic of Korea

    Sang-Je Park, Jae-Won Huh, Young-Hyun Kim, Sang-Rae Lee, Sang-Hyun Kim, Sun-Uk Kim & Kyu-Tae Chang

  2. Department of Biological Sciences, College of Natural Sciences, Pusan National University, Busan, 609-735, Republic of Korea

    Sang-Je Park & Heui-Soo Kim

  3. University of Science & Technology, National Primate Research Center, KRIBB, Chungbuk, 363-883, Republic of Korea

    Young-Hyun Kim & Kyu-Tae Chang

  4. Laboratory of Animal Reproduction and Physiology, Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, 305–764, Republic of Korea

    Min Kyu Kim

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  1. Sang-Je Park
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Correspondence to Kyu-Tae Chang.

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The authors Sang-Je Park, Jae-Won Huh, and Young-Hyun Kim contributed equally to this study.

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Park, SJ., Huh, JW., Kim, YH. et al. Selection of Internal Reference Genes for Normalization of Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) Analysis in the Canine Brain and Other Organs. Mol Biotechnol 54, 47–57 (2013). https://doi.org/10.1007/s12033-012-9543-6

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