从感染HIV的患者中分离外周血单核细胞用于单细胞RNA测序和整合分析。
Isolating peripheral blood mononuclear cells from HIV-infected patients for single-cell RNA sequencing and integration analysis.
作者信息
Jia Jie, Zhao Yu, Wang Jian-Hua, Kuang Yi-Qun
机构信息
NHC Key Laboratory of Drug Addiction Medicine, First Affiliated Hospital of Kunming Medical University & Yunnan Key Laboratory of Stem Cell and Regenerative Medicine, Kunming Medical University, Kunming, Yunnan 650032, China; Scientific Research Laboratory Center, First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650032, China.
Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong 510530, China.
出版信息
STAR Protoc. 2023 Apr 13;4(2):102222. doi: 10.1016/j.xpro.2023.102222.
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)..
对外周血单个核细胞（PBMC）进行单细胞RNA测序（scRNA-seq），有助于剖析HIV感染免疫细胞中的转录变化。在此，我们从HIV感染个体中分离PBMC。在计数细胞数量并验证细胞活力后，我们按照10×基因组学方案和Illumina NovaSeq 6000测序平台对PBMC进行scRNA-seq。此外，在过滤原始序列数据并对基因表达进行归一化后，我们分析B细胞亚群的功能和细胞轨迹以及B细胞受体（BCR）库。有关本方案使用和执行的完整详细信息，请参阅Jia等人（2022年）的文章。
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