Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Bellaterra, Barcelona 08193, Spain.
ETH Zurich Flow Cytomtery Core Facility, Zurich 8093, Switzerland.
STAR Protoc. 2021 Feb 12;2(1):100320. doi: 10.1016/j.xpro.2021.100320. eCollection 2021 Mar 19.
The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s). For complete information on the use and execution of this protocol, please refer to Bologna et al. (2018) and de Leone et al. (2020).
植物组织的特性一直阻碍着人们对 RNA 指导过程的时空和亚细胞分布的理解。在这里，我们描述了一种基于 的通用方案，使用流式细胞分选技术从几乎任何植物物种中研究亚细胞 RNA 分布。该方案包括所有必要的控制步骤，以评估核 RNA 纯化的质量。此外，它可以很容易地应用于不同的植物发育阶段、组织、细胞周期阶段、实验生长条件和特定的细胞类型。有关该方案使用和执行的完整信息，请参考 Bologna 等人（2018 年）和 de Leone 等人（2020 年）的文章。
