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Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis
Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows the dissection of transcriptional changes in immune cells with HIV infection. Here, we isolate PBMCs from HIV-infected individuals. After counting the cell number and verifying cell viability, we perform scRN...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10140154/ https://www.ncbi.nlm.nih.gov/pubmed/37060557 http://dx.doi.org/10.1016/j.xpro.2023.102222 |
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author | Jia, Jie Zhao, Yu Wang, Jian-Hua Kuang, Yi-Qun |
author_facet | Jia, Jie Zhao, Yu Wang, Jian-Hua Kuang, Yi-Qun |
author_sort | Jia, Jie |
collection | PubMed |
description | Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows the dissection of transcriptional changes in immune cells with HIV infection. Here, we isolate PBMCs from HIV-infected individuals. After counting the cell number and verifying cell viability, we perform scRNA-seq for PBMCs on the 10× Genomics protocol and the Illumina NovaSeq 6000 sequencing platform. Furthermore, we analyze the function and cellular trajectories of B cell subsets and B cell receptor (BCR) repertoire after filtering raw sequences data and normalizing gene expression. For complete details on the use and execution of this protocol, please refer to Jia et al. (2022).(1) |
format | Online Article Text |
id | pubmed-10140154 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-101401542023-04-29 Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis Jia, Jie Zhao, Yu Wang, Jian-Hua Kuang, Yi-Qun STAR Protoc Protocol Single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) allows the dissection of transcriptional changes in immune cells with HIV infection. Here, we isolate PBMCs from HIV-infected individuals. After counting the cell number and verifying cell viability, we perform scRNA-seq for PBMCs on the 10× Genomics protocol and the Illumina NovaSeq 6000 sequencing platform. Furthermore, we analyze the function and cellular trajectories of B cell subsets and B cell receptor (BCR) repertoire after filtering raw sequences data and normalizing gene expression. For complete details on the use and execution of this protocol, please refer to Jia et al. (2022).(1) Elsevier 2023-04-14 /pmc/articles/PMC10140154/ /pubmed/37060557 http://dx.doi.org/10.1016/j.xpro.2023.102222 Text en © 2023 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 Jia, Jie Zhao, Yu Wang, Jian-Hua Kuang, Yi-Qun Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title | Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title_full | Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title_fullStr | Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title_full_unstemmed | Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title_short | Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis |
title_sort | isolating peripheral blood mononuclear cells from hiv-infected patients for single-cell rna sequencing and integration analysis |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10140154/ https://www.ncbi.nlm.nih.gov/pubmed/37060557 http://dx.doi.org/10.1016/j.xpro.2023.102222 |
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