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Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing

Isolation of nuclei tagged in specific cell types (INTACT) allows for stress-free and high-throughput analyses of cellular subpopulations. Here, we present an improved protocol for isolation of pure and high-quality GFP-labeled nuclei from frozen livers of INTACT mice, as well as protocols for downs...

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Detalles Bibliográficos
Autores principales: Loft, Anne, Herzig, Stephan, Schmidt, Søren Fisker
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433282/
https://www.ncbi.nlm.nih.gov/pubmed/34527961
http://dx.doi.org/10.1016/j.xpro.2021.100805
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author Loft, Anne
Herzig, Stephan
Schmidt, Søren Fisker
author_facet Loft, Anne
Herzig, Stephan
Schmidt, Søren Fisker
author_sort Loft, Anne
collection PubMed
description Isolation of nuclei tagged in specific cell types (INTACT) allows for stress-free and high-throughput analyses of cellular subpopulations. Here, we present an improved protocol for isolation of pure and high-quality GFP-labeled nuclei from frozen livers of INTACT mice, as well as protocols for downstream sequencing analyses. The adaptation to frozen tissue provides a pause point that allows sampling at multiple time points and/or phenotypic characterization of livers prior to nuclei isolation and downstream analyses. For complete details on the use of this protocol, please refer to Loft et al. (2021).
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spelling pubmed-84332822021-09-14 Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing Loft, Anne Herzig, Stephan Schmidt, Søren Fisker STAR Protoc Protocol Isolation of nuclei tagged in specific cell types (INTACT) allows for stress-free and high-throughput analyses of cellular subpopulations. Here, we present an improved protocol for isolation of pure and high-quality GFP-labeled nuclei from frozen livers of INTACT mice, as well as protocols for downstream sequencing analyses. The adaptation to frozen tissue provides a pause point that allows sampling at multiple time points and/or phenotypic characterization of livers prior to nuclei isolation and downstream analyses. For complete details on the use of this protocol, please refer to Loft et al. (2021). Elsevier 2021-09-08 /pmc/articles/PMC8433282/ /pubmed/34527961 http://dx.doi.org/10.1016/j.xpro.2021.100805 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Loft, Anne
Herzig, Stephan
Schmidt, Søren Fisker
Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title_full Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title_fullStr Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title_full_unstemmed Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title_short Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing
title_sort purification of gfp-tagged nuclei from frozen livers of intact mice for rna- and atac-sequencing
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433282/
https://www.ncbi.nlm.nih.gov/pubmed/34527961
http://dx.doi.org/10.1016/j.xpro.2021.100805
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