Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA 98109, USA.
Department of Immunology, University of Washington, Seattle, WA 98195, USA.
STAR Protoc. 2020 Sep 18;1(2). doi: 10.1016/j.xpro.2020.100092. Epub 2020 Aug 25.
By including oligonucleotide-labeled antibodies into high-throughput single-cell RNA-sequencing protocols, combined transcript and protein expression data can be acquired on the single-cell level. Here, we describe a protocol for the combined analysis of over 40 proteins and 400 genes on over 10 cells using the nano-well based Rhapsody platform. We also include a workflow for sample multiplexing, which uniquely identifies the initial source of cells (such as tissue type or donor) in the downstream analysis after upstream pooling. For complete information on the use and execution of this protocol, please refer to Mair et al. (2020).
通过将寡核苷酸标记的抗体纳入高通量单细胞 RNA 测序方案中,可以在单细胞水平上同时获取转录本和蛋白质表达数据。在这里,我们描述了一种使用基于纳米孔的 Rhapsody 平台对超过 10 个细胞中的 40 多种蛋白质和 400 多个基因进行联合分析的方案。我们还包括了一个样品多路复用的工作流程,该流程在进行上游池化后,在下游分析中唯一地识别细胞的初始来源(如组织类型或供体)。有关此方案的使用和执行的完整信息,请参考 Mair 等人(2020 年)。