Université de Paris, Institut Cochin, INSERM, CNRS, 75014 Paris, France.
Univ Paris Est Créteil, INSERM, IMRB, 94010 Créteil, France.
STAR Protoc. 2021 Jul 28;2(3):100694. doi: 10.1016/j.xpro.2021.100694. eCollection 2021 Sep 17.
Single-nucleus RNA sequencing allows the profiling of gene expression in isolated nuclei. Here, we describe a step-by-step protocol optimized for adult mouse skeletal muscles. This protocol provides two main advantages compared to the widely used single-cell protocol. First, it allows us to sequence the myonuclei of the multinucleated myofibers. Second, it circumvents the cell-dissociation-induced transcriptional modifications. For complete details on the use and execution of this protocol, please refer to Dos Santos et al. (2020) and Machado, Geara et al. (2021).
单细胞 RNA 测序可实现分离核中基因表达的分析。在这里，我们描述了针对成年小鼠骨骼肌进行了优化的分步协议。与广泛使用的单细胞方案相比，该方案具有两个主要优势。首先，它允许我们对多核肌纤维的肌核进行测序。其次，它避免了细胞解离引起的转录修饰。有关此协议的使用和执行的完整详细信息，请参见 Dos Santos 等人（2020 年）和 Machado、Geara 等人（2021 年）。
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