Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and

被引:2
作者
Jia, Jie [1 ,2 ,3 ]
Zhao, Yu [1 ,2 ,3 ]
Wang, Jian-Hua [4 ]
Kuang, Yi-Qun [1 ,2 ,3 ]
机构
[1] Kunming Med Univ, Affiliated Hosp 1, NHC Key Lab Drug Addict Med, Kunming 650032, Yunnan, Peoples R China
[2] Kunming Med Univ, Yunnan Key Lab Stem Cell & Regenerat Med, Kunming 650032, Yunnan, Peoples R China
[3] Kunming Med Univ, Affiliated Hosp 1, Sci Res Lab Ctr, Kunming 650032, Yunnan, Peoples R China
[4] Chinese Acad Sci, Guangzhou Inst Biomed & Hlth, Guangzhou 510530, Guangdong, Peoples R China
来源
STAR PROTOCOLS | 2023年 / 4卷 / 02期
基金
中国国家自然科学基金;
关键词
Bioinformatics; Cell Biology; Cell Isolation; Cell-based Assays; Immunology; Molecular Biology; RNAseq; Sequence Analysis; Single Cell;
D O I
10.1016/j.xpro.2023.102222
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 103 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).
引用
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页数:22
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