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Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq.
Golan, Noa; Cafferty, William B.
Afiliación
  • Golan N; Department of Neurology, Yale University School of Medicine, New Haven, CT 06520, USA.
  • Cafferty WB; Interdepartmental Neuroscience Program, Yale University, New Haven, CT 06520, USA.
STAR Protoc ; 2(4): 100941, 2021 12 17.
Article en En | MEDLINE | ID: mdl-34877546
This protocol provides an improved pipeline for dissociating intact projection neurons from adult mouse cortex for applications including droplet and plate-based single-cell RNA sequencing, qPCR, immunocytochemistry, and long-term in vitro cell culture. This protocol provides a robust and reproducible dissociation pipeline that uses exclusively off-the-shelf reagents, not requiring the use of expensive dissociation kits. The unique incubation steps, in combination with the FACS gating strategy, results in unparalleled enrichment for intact cortical neurons from the adult brain. For complete details on the use and execution of this protocol, please refer to Golan et al. (2021).
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Corteza Cerebral / Neuritas / Análisis de Secuencia de ARN / Análisis de la Célula Individual / Neuronas Límite: Animals Idioma: En Revista: STAR Protoc Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Corteza Cerebral / Neuritas / Análisis de Secuencia de ARN / Análisis de la Célula Individual / Neuronas Límite: Animals Idioma: En Revista: STAR Protoc Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos