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Dissociation of microdissected mouse brain tissue for artifact free single-cell RNA sequencing.

Authors
  • Liu, Lu1
  • Besson-Girard, Simon1
  • Ji, Hao1
  • Gehring, Katrin1
  • Bulut, Buket1
  • Kaya, Tuğberk1, 2, 3
  • Usifo, Fumere1
  • Simons, Mikael4, 2, 3
  • Gokce, Ozgun1, 4
  • 1 Institute for Stroke and Dementia Research, University Hospital, Ludwig-Maximilian-University LMU, 81377 Munich, Germany. , (Germany)
  • 2 Institute of Neuronal Cell Biology, Technical University Munich, 80802 Munich, Germany. , (Germany)
  • 3 German Center for Neurodegenerative Diseases (DZNE), 81377 Munich, Germany. , (Germany)
  • 4 Munich Cluster of Systems Neurology (SyNergy), 81377 Munich, Germany. , (Germany)
Type
Published Article
Journal
STAR protocols
Publication Date
Jun 18, 2021
Volume
2
Issue
2
Pages
100590–100590
Identifiers
DOI: 10.1016/j.xpro.2021.100590
PMID: 34159323
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Single-cell RNA sequencing (scRNA-seq) provides the transcriptome of individual cells and addresses previously intractable problems including the central nervous system's transcriptional responses during health and disease. However, dissociating brain cells is challenging and induces artificial transcriptional responses. Here, we describe an enzymatic dissociation method for mouse brain that prevents dissociation artifacts and lowers technical variations with standardized steps. We tested this protocol on microdissected brain tissue of 3-week- to 24-month-old mice and obtained high-quality scRNA-seq results. For complete details on the use and execution of this protocol, please refer to Safaiyan et al. (2021). © 2021 The Author(s).

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