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Nuclei isolation from surgically resected human hippocampus

Fatma Ayhan, Connor Douglas, Bradley Lega, Geneviève Konopka

2021STAR Protocols27 citationsDOIOpen Access PDF

Abstract

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).

Topics & Concepts

TranscriptomeBiologyHuman brainHippocampusRNAComputational biologyIsolation (microbiology)Brain tissueCell biologyNeuroscienceBioinformaticsGeneticsGene expressionGeneSingle-cell and spatial transcriptomicsMolecular Biology Techniques and ApplicationsCell Image Analysis Techniques
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