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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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). |
format | Online Article Text |
id | pubmed-8433282 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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