Highly parallel and efficient single cell mRNA sequencing with paired picoliter chambers

Posted: 2020-05-08   Visits: 42

Authors: Mingxia Zhang, Yuan Zou, Xing Xu, Xuebing Zhang, Mingxuan Gao, Jia Song, Peifeng Huang, Qin Chen, Zhi Zhu, Wei Lin, Richard N. Zare, Chaoyong Yang

Abstract: ScRNA-seq has the ability to reveal accurate and precise cell types and states. Existing scRNA-seq platforms utilize bead-based technologies uniquely barcoding individual cells, facing practical challenges for precious samples with limited cell number. Here, we present a scRNA-seq platform, named Paired-seq, with high cells/beads utilization efficiency, cell-free RNAs removal capability, high gene detection ability and low cost. We utilize the differential flow resistance principle to achieve single cell/barcoded bead pairing with high cell utilization efficiency (95%). The integration of valves and pumps enables the complete removal of cell-free RNAs, efficient cell lysis and mRNA capture, achieving highest mRNA detection accuracy (R = 0.955) and comparable sensitivity. Lower reaction volume and higher mRNA capture and barcoding efficiency significantly reduce the cost of reagents and sequencing. The single-cell expression profile of mES and drug treated cells reveal cell heterogeneity, demonstrating the enormous potential of Paired-seq for cell biology, developmental biology and precision medicine.

Link: https://www.nature.com/articles/s41467-020-15765-0