TAS-Seq: a robust and sensitive amplification method for bead-based scRNA-seq

Abstract

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. We developed terminator-assisted solid-phase cDNA amplification and sequencing (TAS-Seq), a scRNA-seq method relying on a terminator, terminal transferase, and nanowell/beads-based scRNA-seq platform that could acquire scRNA-seq data, is highly correlated with flow-cytometric data, has gene-detection sensitivity, and is more robust than widely-used methods.

Single-Cell Analysis Revealed the Role of CD8+ Effector T Cells in Preventing Cardioprotective Macrophage Differentiation in the Early Phase of Heart Failure

“…One week after TAC surgery, heart CD45⁺ cells were isolated using an autoMACS Pro Separator by positive selection after Percoll enrichment. To obtain single-cell RNA-seq data, we used an BD Rhapsody system (BD Biosciences) and TAS-Seq as previously reported ( 40 ). A single-cell suspension of ~20,000 cells was captured from all isolated cells, without selection, on an array of >200,000 microwells through a limited dilution approach.…”

CD8+T-cell memory induced by successive SARS-CoV-2 mRNA vaccinations is characterized by clonal replacement

“…Data processing of scRNAseq reads was performed as described in Shichino et al ( 59 ). Adapter trimming of sequencing data was performed by using cutadapt 3.4 ( 53 ).…”

Chondrocyte-like cells in nucleus pulposus and articular chondrocytes have similar transcriptomic profiles and are paracrine-regulated by hedgehog from notochordal cells and subchondral bone

“…Library preparation and the first processing of paired-end fastq files were performed according to the workflow in the previously reported TAS-seq method (Shichino et al, 2022). In brief, sorted single-cell suspensions were used to synthesize cDNA with BD Rhapsody Express Single-Cell Analysis System (BD Biosciences) using a BD Rhapsody Targeted and Abseq Reagent kit (BD Biosciences) following the manufacturer's instructions.…”