Chinese Bulletin of Botany ›› 2012, Vol. 47 ›› Issue (4): 427-436.DOI: 10.3724/SP.J.1259.2012.00427

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Characterization and Selection of Reference Genes for Real-time Quantitative RT-PCR of Plants

Wei Yuan1,2, Hongjian Wan1, Yuejian Yang1*   

  1. 1Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

    2College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2011-10-28 Revised:2012-03-19 Online:2012-07-01 Published:2012-07-26
  • Contact: Yuejian Yang

Abstract: Real-time quantitative RT-PCR (qRT-PCR) is one of the most common technologies used for gene expression and transcriptome analysis, with its high sensitivity, specificity, good reproducibility, wide dynamic quantification range and high-throughput capacity. Selecting the appropriate reference genes is the first step in analyzing the expression of genes of interest. Selecting appropriate reference genes depends on experimental conditions, and selection of reference genes changes after the experimental conditions. Therefore, the accuracy of results from qRT-PCR analysis largely depends on the reference genes used. In this paper, we give a comprehensive summary of reference genes for qRT-PCR, including their selection, characteristics of traditional reference genes, mining new reference genes, the advantage of combining different reference genes, and how to assess stable reference gene expression. The results provide a theoretical foundation for selecting appropriate reference genes for qRT-PCR of plants.