single-cell RNA-seq of murine spleen by TAS-Seq. single-cell RNA-seq of murine spleen by TAS-Seq

Single-cell RNA-sequencing (scRNA-seq) is valuable for analyzing cellular heterogeneity. Cell composition accuracy is critical for analyzing cell-cell interaction networks from scRNA-seq data because cell abundancy might affect to the network. We developed terminator-assisted solid-phase cDNA amplification and sequencing (TAS-Seq), a scRNA-seq method that relied on terminator, terminal transferase, and nanowell/beads-based scRNA-seq platform, that could acquire scRNA-seq data with higher correlation with flow-cytometric data, gene-detection sensitivity, and robustness than widely-used methods. Overall design: To clarify performance of TAS-Seq, we analyzed mouse spleen cells collected from subcutaneous lewis lung carcinoma (LLC) model. 5x105 LLC cells were subcutaneously injected to 10 week old C57BL/6J female mice, treated with 200 μg/shot anti-CD4 antibody at on day 5 and day 9 post injection, and spleen were recovered at day 12. Resulted spleen cells were subjected by BD Rhapsody workflow, and half of BD Rhapsody beads were processed by TAS-Seq. and the other half of the beads were processed by BD official WTA protocol.