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优化鼻腔黏膜样本处理以获取高质量 scRNA-seq 和 scATAC-seq 数据的方案。

Protocol for optimized nasal mucosa sample processing to obtain high-quality scRNA-seq and scATAC-seq data.

机构信息

BGI Research, Shenzhen 518083, China.

BGI Research, Shenzhen 518083, China; Department of Otolaryngology, Head and Neck Surgery, Second Hospital, Shanxi Medical University, Taiyuan 030001, China; Shanxi Medical University-BGI Collaborative Center for Future Medicine, Shanxi Medical University, Taiyuan 030001, China.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103298. doi: 10.1016/j.xpro.2024.103298. Epub 2024 Sep 7.

Abstract

Examining nasal mucosa samples is crucial for nasal cavity disease research and diagnosis. Simultaneously obtaining high-quality data for single-cell transcriptomics (single-cell RNA sequencing [scRNA-seq]) and epigenomics (single-cell assay for transposase-accessible chromatin using sequencing [scATAC-seq]) of nasal mucosa tissues is challenging. Here, we present a protocol for processing human nasal mucosa samples to obtain data for both scRNA-seq and scATAC-seq. We describe steps for extracting human nasal mucosa tissue, mechanical and enzymatic dissociation, lysis of red blood cells, and a viability assay. We then detail procedures for library preparation and quality control.

摘要

检查鼻腔黏膜样本对于鼻腔疾病的研究和诊断至关重要。同时获得高质量的单细胞转录组学(单细胞 RNA 测序 [scRNA-seq])和表观基因组学(使用测序进行转座酶可及染色质的单细胞分析 [scATAC-seq])数据对于鼻腔黏膜组织具有挑战性。在这里,我们提出了一种处理人鼻腔黏膜样本以获取 scRNA-seq 和 scATAC-seq 数据的方案。我们描述了提取人鼻腔黏膜组织、机械和酶解、红细胞裂解以及活力测定的步骤。然后,我们详细介绍了文库制备和质量控制的程序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/886a/11407050/6d6723e7ba43/fx1.jpg

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