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首页> 美国卫生研究院文献>BMC Plant Biology >Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process
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Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process

机译：番茄发育过程中用于实时定量RT-PCR研究的内部控制基因的选择

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摘要

BackgroundThe elucidation of gene expression patterns leads to a better understanding of biological processes. Real-time quantitative RT-PCR has become the standard method for in-depth studies of gene expression. A biologically meaningful reporting of target mRNA quantities requires accurate and reliable normalization in order to identify real gene-specific variation. The purpose of normalization is to control several variables such as different amounts and quality of starting material, variable enzymatic efficiencies of retrotranscription from RNA to cDNA, or differences between tissues or cells in overall transcriptional activity. The validity of a housekeeping gene as endogenous control relies on the stability of its expression level across the sample panel being analysed. In the present report we describe the first systematic evaluation of potential internal controls during tomato development process to identify which are the most reliable for transcript quantification by real-time RT-PCR.
机译：背景技术对基因表达模式的阐明使人们对生物学过程有了更好的了解。实时定量RT-PCR已成为深入研究基因表达的标准方法。对靶mRNA量具有生物学意义的报告需要准确可靠的归一化，以鉴定真正的基因特异性变异。标准化的目的是控制几个变量，例如起始材料的数量和质量不同，从RNA到cDNA的逆转录的可变酶促效率，或组织或细胞之间在总体转录活性方面的差异。管家基因作为内源性对照的有效性取决于其在整个分析样本中的表达水平的稳定性。在本报告中，我们描述了对番茄发育过程中潜在内部控制的第一个系统评价，以确定哪个是实时RT-PCR转录定量最可靠的方法。

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