优化的单分子 RNA FISH 方案，用于可视化 中的基因表达。
Optimized protocol for single-molecule RNA FISH to visualize gene expression in .
机构信息
Division of Gene Regulation, The Netherlands Cancer Institute, Oncode Institute, 1066CX Amsterdam, the Netherlands.
出版信息
STAR Protoc. 2021 Jul 6;2(3):100647. doi: 10.1016/j.xpro.2021.100647. eCollection 2021 Sep 17.
Single-molecule RNA fluorescence hybridization (smFISH) allows subcellular visualization, localization, and quantification of endogenous RNA molecules in fixed cells. The spatial and intensity information of each RNA can be used to distinguish mature from nascent transcripts inside each cell, revealing both past and instantaneous transcriptional activity. Here, we describe an optimized protocol for smFISH in with optimized lyticase digestion time and hybrization steps for more homogenous results. For complete details on the use and execution of this protocol, please refer to Donovan et al. (2019).
单分子 RNA 荧光杂交 (smFISH) 允许在固定细胞中对内源性 RNA 分子进行亚细胞可视化、定位和定量。每个 RNA 的空间和强度信息可用于区分每个细胞内成熟和新生转录本，从而揭示过去和瞬时的转录活性。在这里，我们描述了一个在 中优化的 smFISH 方案，包括优化的溶酶酶消化时间和杂交步骤，以获得更均匀的结果。有关该方案的使用和执行的完整详细信息，请参阅 Donovan 等人。(2019)。
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