Profiling low-mRNA content cells in complex human tissues using BD Rhapsody single-cell analysis

The successful recovery of immune cells, particularly those with low mRNA content, by single-cell RNA sequencing (scRNA-seq) remains a significant challenge. Tissue dissociation and selection of the appropriate scRNA-seq technology are crucial. Our protocol efficiently recovers low-mRNA content immune cells using the BD Rhapsody scRNA-seq platform. It includes optimized tissue dissociation for prostate, lung, and liver tissues, cell labeling with Sample Tag antibodies, microwell-based single-cell capture, cDNA synthesis, library preparation, and data pre-processing with basic quality control analysis. For complete details on the use and execution of this protocol, please refer to Salcher et al.1. Keywords: Bioinformatics; Cancer; RNAseq; Sequencing.