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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...

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Detalles Bibliográficos
Autores principales: Jia, Jie, Zhao, Yu, Wang, Jian-Hua, Kuang, Yi-Qun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
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)
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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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