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所在平台: Udemy |
课程主页: https://www.udemy.com/course/principles-in-real-time-pcr-and-data-analysis/
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**课程名称：**qPCR与数据分析原理 (Principles in qPCR and Data analysis) **课程概述：** 本课程深入探讨了定量聚合酶链式反应 (qPCR)，一种能够实时测定PCR产物量的强大技术，尤其在基因表达研究中用途广泛。课程将区分qPCR、实时PCR以及可能引起混淆的RT-PCR（根据应用情况，可能是定量逆转录PCR）。 **核心优势：** * **高精度和灵敏度：** qPCR相比传统PCR，能够以高精度和高灵敏度，在一个宽广的动态范围内确定模板DNA（扩增靶序列）的初始拷贝数。 * **结果多样性：** qPCR结果可以是定性（序列的有无）或定量的（拷贝数）。 **应用领域：** * **基因拷贝数测定：** 在研究实验室中，qPCR广泛用于定量测量转化细胞系中的基因拷贝数（基因剂量）或突变基因的存在。 * **基因表达量化：** 结合逆转录PCR (RT-PCR)，qPCR能够精确量化基因表达的变化，例如，通过测量细胞mRNA水平的变化，来评估在不同环境条件或药物处理下表达的升高或降低。 **课程内容亮点：** * **定义与概念：** 明确qPCR的定义和基本概念。 * **DNA检测方法：** 介绍直接使用DNA结合染料和间接使用特异性荧光标记物进行DNA检测的原理。 * **实验方法选择：** 指导学员如何为不同实验选择合适的qPCR方法。 * **qPCR程序设置：** 讲解如何执行qPCR实验程序。 * **故障排除：** 提供qPCR实验中常见的故障排除技巧。 * **数据分析：** 重点讲解qPCR数据的分析方法。
Quantitative polymerase chain reaction (Q-PCR) is a method by which the amount of the PCR product can be determined, in real-time, and is very useful for investigating gene expression. The qPCR method is sometimes also referred to as real-time PCR or depending on the application, quantitative reverse-transcriptase PCR (both of which are abbreviated to RT-PCR, which can be rather confusing). The main advantage of real-time PCR over PCR is that real-time PCR allows you to determine the initial number of copies of template DNA (the amplification target sequence) with accuracy and high sensitivity over a wide dynamic range. Real-time PCR results can either be qualitative (the presence or absence of a sequence) or quantitative (copy number). Quantitative real-time PCR is thus also known as qPCR analysis. In research laboratories, qPCR assays are widely used for the quantitative measurement of gene copy number (gene dosage) in transformed cell lines or the presence of mutant genes. In combination with reverse-transcription PCR (RT-PCR), qPCR assays can be used to precisely quantitate changes in gene expression, for example, an increase or decrease in expression in response to different environmental conditions or drug treatment, by measuring changes in cellular mRNA levels. This course is on real-time PCR, definition, concept, and types of DNA detection directly using DNA binding dye or indirectly using specific fluorescent dye. What is the suitable method for each experiment? How can we perform the qPCR program? Troubleshooting in qPCR. Data analysis of qPCR output.
