Liu Shang-Yun, Rink Jochen C
Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
Methods Mol Biol. 2018;1774:259-265. doi: 10.1007/978-1-4939-7802-1_6.
Isolation of high-quality and nondegraded RNA is a prerequisite for many modern molecular biology applications. A variety of RNA extraction products and protocols are available for the standard model organisms, yet species-specific peculiarities of less well studied organisms often require specific protocol adaptations. Here we describe a robust RNA extraction protocol for planarians that is widely used in the community. The protocol combines tissue homogenization in TRIzol with phenol-chloroform extraction and subsequent purification over commercial columns. The purified RNA can be used for many downstream applications, such as cDNA synthesis and next-generation sequencing (NGS, RNA-seq).
分离高质量且未降解的RNA是许多现代分子生物学应用的前提条件。对于标准模式生物,有多种RNA提取产品和方案可供选择,然而,对于研究较少的生物,其物种特异性特点往往需要对方案进行特定调整。在此,我们描述了一种在该领域广泛使用的用于涡虫的可靠RNA提取方案。该方案将在TRIzol中进行组织匀浆与酚-氯仿提取相结合,随后通过商业柱进行纯化。纯化后的RNA可用于许多下游应用,如cDNA合成和下一代测序(NGS,RNA测序)。