Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq

Summary: 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 r...

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Autores principales:	Noa Golan, William B. Cafferty
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Cell Biology Cell isolation Single Cell Flow Cytometry/Mass Cytometry RNAseq Neuroscience Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/dc0fab82d2094529a55604614fe134ca
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spelling	oai:doaj.org-article:dc0fab82d2094529a55604614fe134ca2021-11-26T04:40:31ZDissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq2666-166710.1016/j.xpro.2021.100941https://doaj.org/article/dc0fab82d2094529a55604614fe134ca2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S266616672100647Xhttps://doaj.org/toc/2666-1667Summary: 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).Noa GolanWilliam B. CaffertyElsevierarticleCell BiologyCell isolationSingle CellFlow Cytometry/Mass CytometryRNAseqNeuroscienceScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100941- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cell Biology Cell isolation Single Cell Flow Cytometry/Mass Cytometry RNAseq Neuroscience Science (General) Q1-390
spellingShingle	Cell Biology Cell isolation Single Cell Flow Cytometry/Mass Cytometry RNAseq Neuroscience Science (General) Q1-390 Noa Golan William B. Cafferty Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
description	Summary: 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).
format	article
author	Noa Golan William B. Cafferty
author_facet	Noa Golan William B. Cafferty
author_sort	Noa Golan
title	Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
title_short	Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
title_full	Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
title_fullStr	Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
title_full_unstemmed	Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq
title_sort	dissociation of intact adult mouse cortical projection neurons for single-cell rna-seq
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/dc0fab82d2094529a55604614fe134ca
work_keys_str_mv	AT noagolan dissociationofintactadultmousecorticalprojectionneuronsforsinglecellrnaseq AT williambcafferty dissociationofintactadultmousecorticalprojectionneuronsforsinglecellrnaseq
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Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq

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