Real-Time Quantitative PCR: Primer Design, Reference Gene Selection, Calculations and Statistics

文献类型： 外文期刊

第一作者： Chen, Zhiwei

作者： Chen, Zhiwei;Liu, Chenghong;Halford, Nigel G.

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关键词： quantitative real-time PCR; relative gene expression; normalized relative quantity; PCR efficiency

期刊名称：METABOLITES （ 影响因子：4.1； 五年影响因子：4.5 ）

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年卷期： 2023 年 13 卷 7 期

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收录情况： SCI

摘要： Real-time quantitative PCR is a technique that can measure the content of the target nucleic acid sequence of interest in a given sample. It is mainly divided into absolute and relative quantitative methods. The relative quantification is mainly used in gene expressions for functional genomic and transcriptome studies. However, to use this technology accurately, there are some key points to master. First, specific primers need to be designed to ensure amplification of the gene of interest (GOI). Second, the appropriate reference gene or reference gene combination has to be selected. Finally, scientific gene expression level calculations and statistics are required to obtain accurate results. Therefore, this work proposes a workflow for relative quantitative PCR and introduces the relevant points so that beginners can better understand and use this technology.

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