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A protocol for single nucleus RNA-seq from frozen skeletal muscle

Single-cell technologies are a method of choice to obtain vast amounts of cell-specific transcriptional information under physiological and diseased states. Myogenic cells are resistant to single-cell RNA sequencing because of their large, multinucleated nature. Here, we report a novel, reliable, an...

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Autores principales: Soule, Tyler GB, Pontifex, Carly S, Rosin, Nicole, Joel, Matthew M, Lee, Sukyoung, Nguyen, Minh Dang, Chhibber, Sameer, Pfeffer, Gerald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Life Science Alliance LLC 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10011611/
https://www.ncbi.nlm.nih.gov/pubmed/36914268
http://dx.doi.org/10.26508/lsa.202201806
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author Soule, Tyler GB
Pontifex, Carly S
Rosin, Nicole
Joel, Matthew M
Lee, Sukyoung
Nguyen, Minh Dang
Chhibber, Sameer
Pfeffer, Gerald
author_facet Soule, Tyler GB
Pontifex, Carly S
Rosin, Nicole
Joel, Matthew M
Lee, Sukyoung
Nguyen, Minh Dang
Chhibber, Sameer
Pfeffer, Gerald
author_sort Soule, Tyler GB
collection PubMed
description Single-cell technologies are a method of choice to obtain vast amounts of cell-specific transcriptional information under physiological and diseased states. Myogenic cells are resistant to single-cell RNA sequencing because of their large, multinucleated nature. Here, we report a novel, reliable, and cost-effective method to analyze frozen human skeletal muscle by single-nucleus RNA sequencing. This method yields all expected cell types for human skeletal muscle and works on tissue frozen for long periods of time and with significant pathological changes. Our method is ideal for studying banked samples with the intention of studying human muscle disease.
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spelling pubmed-100116112023-03-15 A protocol for single nucleus RNA-seq from frozen skeletal muscle Soule, Tyler GB Pontifex, Carly S Rosin, Nicole Joel, Matthew M Lee, Sukyoung Nguyen, Minh Dang Chhibber, Sameer Pfeffer, Gerald Life Sci Alliance Methods Single-cell technologies are a method of choice to obtain vast amounts of cell-specific transcriptional information under physiological and diseased states. Myogenic cells are resistant to single-cell RNA sequencing because of their large, multinucleated nature. Here, we report a novel, reliable, and cost-effective method to analyze frozen human skeletal muscle by single-nucleus RNA sequencing. This method yields all expected cell types for human skeletal muscle and works on tissue frozen for long periods of time and with significant pathological changes. Our method is ideal for studying banked samples with the intention of studying human muscle disease. Life Science Alliance LLC 2023-03-13 /pmc/articles/PMC10011611/ /pubmed/36914268 http://dx.doi.org/10.26508/lsa.202201806 Text en © 2023 Soule et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).
spellingShingle Methods
Soule, Tyler GB
Pontifex, Carly S
Rosin, Nicole
Joel, Matthew M
Lee, Sukyoung
Nguyen, Minh Dang
Chhibber, Sameer
Pfeffer, Gerald
A protocol for single nucleus RNA-seq from frozen skeletal muscle
title A protocol for single nucleus RNA-seq from frozen skeletal muscle
title_full A protocol for single nucleus RNA-seq from frozen skeletal muscle
title_fullStr A protocol for single nucleus RNA-seq from frozen skeletal muscle
title_full_unstemmed A protocol for single nucleus RNA-seq from frozen skeletal muscle
title_short A protocol for single nucleus RNA-seq from frozen skeletal muscle
title_sort protocol for single nucleus rna-seq from frozen skeletal muscle
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10011611/
https://www.ncbi.nlm.nih.gov/pubmed/36914268
http://dx.doi.org/10.26508/lsa.202201806
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