Affiliation:
1. Laboratory of Molecular and Behavioral Neuroscience, Institute for Neuroscience, Department of Health Sciences and Technology ETH Zurich Zurich Switzerland
2. Neuroscience Center Zurich ETH Zurich and University of Zurich Zurich Switzerland
Abstract
AbstractNeuroscience research greatly benefits from single‐cell sequencing technologies, which can reveal transcriptional alterations on a cellular level. However, preparing single‐cell suspensions is technically challenging, requires experience, and has several limitations that can influence the transcriptional readout. Performing sequencing of single nuclei instead of single cells alleviates several of the challenges of sample preparation and highlights acute nuclear transcription. Here, we provide a protocol to prepare a nuclei suspension for single‐nucleus RNA‐sequencing for cell type‐specific transcriptional profiling of brain tissue using the 10x Genomics single‐cell gene expression assay. Furthermore, we highlight important aspects to consider during experimental design and data analysis. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Preparation of single‐nucleus suspensionBasic Protocol 2: Preparation and sequencing of single‐nucleus libraries for RNA‐seq
Subject
Medical Laboratory Technology,Health Informatics,General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Neuroscience
Cited by
3 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献