Standard and targeted single-cell transcriptomics of CD8+ T cell samples and targeted single-cell transcriptomics of B cell samples for systematic analysis

We developed a straightforward targeted sequencing approach as extension to high-throughput droplet-based scRNA-seq. By overlaying standard gene expression data with genotype information at the transcript level, we created a powerful method for evaluating the impact of genetic variants. We systematically analysed the effect of technical parameters on the targeted sequencing method and demonstrated how the targeted sequencing method can be used. T, CD8+ T or B lymphocytes were isolated from PBMCs using EasySep Human T, CD8+ T or B Cell Isolation Kits (STEMCELL Technologies). Single cell suspensions were used for single cell transcriptomics with the Chromium Next GEM Single Cell 3’ Reagent Kits v3.1 (10x Genomics; CD8+ T and B cells) or the Chromium Single Cell V(D)J Reagent Kits v.1.1 (10x Genomics; T cells). Left-over amplified cDNA was used as input for our targeted sequencing extension. -------------------------- Authors state "Raw data files cannot be shared because of GDPR."