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Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis
Current single-cell RNA sequencing (scRNA-seq) protocols are limited by the number of cells that can be simultaneously sequenced, restricting the ability to resolve heterogeneity of rare cell types. We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellu...
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/PMC8605103/ https://www.ncbi.nlm.nih.gov/pubmed/34825218 http://dx.doi.org/10.1016/j.xpro.2021.100957 |
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author | Dang, Mai T. Mafra, Fernanda Haldar, Malay |
author_facet | Dang, Mai T. Mafra, Fernanda Haldar, Malay |
author_sort | Dang, Mai T. |
collection | PubMed |
description | Current single-cell RNA sequencing (scRNA-seq) protocols are limited by the number of cells that can be simultaneously sequenced, restricting the ability to resolve heterogeneity of rare cell types. We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellum without cell sorting to minimize cell damage for scRNA-seq. This protocol includes the procedures for further enrichment of myeloid cells using CD11b+ magnetic beads, followed by the generation of scRNA library and sequencing analysis. For complete details on the use and execution of this protocol, please refer to Dang et al. (2021). |
format | Online Article Text |
id | pubmed-8605103 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-86051032021-11-24 Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis Dang, Mai T. Mafra, Fernanda Haldar, Malay STAR Protoc Protocol Current single-cell RNA sequencing (scRNA-seq) protocols are limited by the number of cells that can be simultaneously sequenced, restricting the ability to resolve heterogeneity of rare cell types. We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellum without cell sorting to minimize cell damage for scRNA-seq. This protocol includes the procedures for further enrichment of myeloid cells using CD11b+ magnetic beads, followed by the generation of scRNA library and sequencing analysis. For complete details on the use and execution of this protocol, please refer to Dang et al. (2021). Elsevier 2021-11-17 /pmc/articles/PMC8605103/ /pubmed/34825218 http://dx.doi.org/10.1016/j.xpro.2021.100957 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 Dang, Mai T. Mafra, Fernanda Haldar, Malay Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title | Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title_full | Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title_fullStr | Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title_full_unstemmed | Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title_short | Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis |
title_sort | isolation of myeloid cells from mouse brain tumors for single-cell rna-seq analysis |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8605103/ https://www.ncbi.nlm.nih.gov/pubmed/34825218 http://dx.doi.org/10.1016/j.xpro.2021.100957 |
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