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

STAR Protocols

Simon Besson-Girard¹  Ozgun Gokce²  Mikael Simons³  Lu Liu⁴  Buket Bulut⁴  Tuğberk Kaya⁴  Katrin Gehring⁴  Fumere Usifo⁴  Hao Ji⁴ 

[1] Corresponding author;German Center for Neurodegenerative Diseases (DZNE), 81377 Munich, Germany;Institute of Neuronal Cell Biology, Technical University Munich, 80802 Munich, Germany;Institute for Stroke and Dementia Research, University Hospital, Ludwig-Maximilian-University LMU, 81377 Munich, Germany;

关键词: Cell Biology;    Cell isolation;    Single Cell;    Flow Cytometry/Mass Cytometry;    Sequencing;    RNA-seq;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown