School of Biological Science, Luoyang Normal University, Luoyang, China.
Methods Mol Biol. 2021;2326:95-103. doi: 10.1007/978-1-0716-1514-0_7.
Semi-quantitative reverse transcription and polymerase chain reaction (sqRT-PCR) is a simple and specific method for quantitative RNA in recent years. The relative quantity of a specific mRNA in the samples can be inferred by reverse transcription of mRNA into cDNA, and PCR amplification and determination of the quantity of PCR products. The semi-quantitative analysis is carried out under a fixed number of PCR cycles, and the total RNA concentration is kept in the exponential phase of the PCR. The method is to use a housekeeping gene (usually actin, GAPDH, and EF1α) as a reference standard in treated and control organisms to observe the expression of the interested genes (upregulated or downregulated) in toxicology. In this chapter, we describe a step-by-step method for determining the differential regulation of target genes in organisms exposed to environmental pollutants.
近年来，半定量逆转录聚合酶链反应（sqRT-PCR）是一种用于定量 RNA 的简单而特异的方法。通过将 mRNA 逆转录成 cDNA，并进行 PCR 扩增和 PCR 产物定量，可以推断出样品中特定 mRNA 的相对含量。半定量分析是在固定的 PCR 循环数下进行的，总 RNA 浓度保持在 PCR 的指数期。该方法是在处理和对照生物中使用管家基因（通常为肌动蛋白、GAPDH 和 EF1α）作为参考标准，以观察毒物暴露后感兴趣基因（上调或下调）的表达。在本章中，我们描述了一种用于确定暴露于环境污染物的生物体中靶基因差异调控的分步方法。
