Translational Immunology Research Program, University of Helsinki, Helsinki, Finland.
Department of Bacteriology and Immunology, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
Methods Mol Biol. 2020;2184:31-45. doi: 10.1007/978-1-0716-0802-9_3.
Transcriptome analysis at a single-cell level with single-cell RNA sequencing (scRNA-seq) is a powerful method for detailed characterization of heterogeneous cell populations. Recent developments have enabled parallel analysis of both transcript and protein levels by using antibodies conjugated to barcoded oligonucleotides. These antibodies enable protein levels to be converted into nucleotide format, allowing the sequencing-based detection of both modalities at single-cell level. Here we present a simple and reliable method for conjugation of oligonucleotides with antibodies and a protocol for their use in single-cell transcriptome sequencing.
单细胞 RNA 测序 (scRNA-seq) 的转录组分析是一种对异质细胞群体进行详细描述的强大方法。最近的发展使通过使用与条形码寡核苷酸偶联的抗体对转录本和蛋白质水平进行平行分析成为可能。这些抗体使蛋白质水平能够转化为核苷酸形式,从而允许在单细胞水平上基于测序检测两种模式。在此,我们提出了一种简单可靠的将寡核苷酸与抗体偶联的方法,以及一种将其用于单细胞转录组测序的方案。