Department of Neuroscience, UT Southwestern Medical Center, Dallas, TX 75390, USA.
Department of Neurosurgery, UT Southwestern Medical Center, Dallas, TX 75390, USA.
STAR Protoc. 2021 Sep 16;2(4):100844. doi: 10.1016/j.xpro.2021.100844. eCollection 2021 Dec 17.
Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).
单细胞 RNA 测序（snRNA-seq），其中核转录组是细胞转录组的代表，已被用于对人类大脑进行分析。snRNA-seq 对组织处理、组织质量、死后间隔时间和细胞碎片敏感。本方案概述了从手术切除的人脑组织中分离高质量核的步骤，然后进行蔗糖梯度分离，得到神经元核和非神经元核，从而能够对各种细胞类型进行无偏分析。有关此方案的使用和执行的完整详细信息，请参阅 Ayhan 等人（2021 年）。
