Benchmarking full-length transcript single cell RNA sequencing protocols

Abstract

Single cell mRNA sequencing technologies have transformed our understanding of cellular heterogeneity and identity. For sensitive discovery or clinical marker estimation where high transcript capture per cell is needed only plate-based techniques currently offer sufficient resolution. Here, we present a performance evaluation of three different plate-based scRNA-seq protocols. Our evaluation is aimed towards applications requiring high gene detection sensitivity, reproducibility between samples, and minimum hands-on time, as is required, for example, in clinical use. We included two commercial kits, NEBNext® Single Cell/ Low Input RNA Library Prep Kit (NEB®), SMART-seq® HT kit (Takara®), and the non-commercial protocol Genome & Transcriptome sequencing (G&T). G&T delivered the highest detection of genes per single cell, at the absolute lowest price. Takara® kit presented similar high gene detection per single cell, and high reproducibility between sample, but at the absolute highest price. NEBNext® delivered a lower detection of genes but remain an alternative to more expensive commercial kits.