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所在平台: Udemy |
课程主页: https://www.udemy.com/course/isolation-of-dna-and-rna-concepts/
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课程名称：DNA和RNA概念的分离 课程概述： 本课程旨在介绍RNA的分离技术，包括样本收集与保护、RNA准备、RNA定量和存储等步骤。首先，样本的选择与处理方法对于确保RNA的产量和质量至关重要。在适当的条件下，4°C下保存的样本即使存放一个月，仍能保持完整的RNA。 在RNA准备阶段，市场上有多种技术可供选择，这些技术可以分为四种主要方法：有机提取法、离心篮格式、磁颗粒法和直接裂解法。虽然这些方法都能得到适用于多种分析技术的高质量RNA，但在选择合适的提取技术时需考虑多个因素。 直接裂解法利用裂解缓冲液进行样本准备，通过破坏样本、稳定核酸，并与后续分析兼容的方式实施。这种方法省去了从固体表面进行结合和洗脱的步骤，避免了可能出现的偏倚和回收效率问题，使得直接裂解法成为一种方便而有效的选择。 通过本课程，学习者将掌握RNA的分离与保存的各个关键步骤，为后续的分析工作奠定基础。
·Introduction:Step 1: Sample Collection and ProtectionStep 2: RNA PreparationStep 3: Quantitation of Isolated RNAStep 4: Storage of Isolated RNAStep 1: Sample Collection and ProtectionFinding the most appropriate method of cell or tissue disruption for your specific starting material is important for maximizing the yield and quality of your RNA preparation. During sample disruption for RNA isolation, Samples stored at 4°C generate intact RNA, even after storage for a month.Step 2: RNA PreparationA number of RNA preparation technologies are widely available that can be classified into four general techniques: organic extraction methods, spin basket formats, magnetic particle methods, and direct lysis methods. While all can be used to prepare high-quality RNA suitable for a wide variety of analysis techniques, there are several factors to consider in selecting the right purification technology.Direct Lysis MethodsDirect lysis methods perform sample preparation (not purification) by utilizing lysis buffer formulations that disrupt samples, stabilize nucleic acids, and are compatible with downstream analysis. Typically, a sample is mixed with lysis agent, incubated for some amount of time under specified conditions, and then used directly for downstream analysis. If desired, samples can often be purified from stabilized lysates. By eliminating the need to bind and elute from solid surfaces, direct lysis methods can avoid bias and recovery efficiency effects that may occur when using other purification methods.
