CEL-Seq2: sensitive highly-multiplexed single-cell RNA-Seq. CEL-Seq2: sensitive highly-multiplexed single-cell RNA-Seq

Single-cell transcriptomics requires a method that is sensitive, accurate, and reproducible. Here, we present CEL-Seq2, a modified version of our CEL-Seq method, with three-fold higher sensitivity, lower costs, and less hands-on time. We also implemented CEL-Seq2 on Fluidigm’s C1 system, thereby providing its first single-cell, on-chip barcoding method, and detected gene expression changes accompanying the progression through the cell cycle in mouse fibroblast cells. We also compare with Smart-Seq to demonstrate CEL-Seq2’s increased sensitivity relative to other available methods. Collectively, the improvements make CEL-Seq2 uniquely suited to single-cell RNA-Seq analysis in terms of economics, resolution, and ease of use Overall design: Single mouse dendritic and fibroblast cells were analysed for comaprison of CEL-Seq2 to CEL-Seq and Smart-Seq on C1. C. elegans totat RNA was used to calibrate and optimze the new CEL-Seq2 protocol.

单细胞转录组学（single-cell transcriptomics）亟需兼具高灵敏度、准确可靠与可重复的研究方法。本文介绍CEL-Seq2——本团队前期开发的CEL-Seq方法的改良版本，其灵敏度较原方法提升三倍，成本更低，且手工操作耗时更短。我们将CEL-Seq2应用于Fluidigm的C1系统，由此推出首款单细胞芯片内条形码标记方法，并在小鼠成纤维细胞中检测到伴随细胞周期进程的基因表达变化。我们还通过与Smart-Seq比对，证实CEL-Seq2相较于现有其他方法具有更优的灵敏度。综上，上述改良使得CEL-Seq2在经济性、分辨率与易用性方面，成为唯一适用于单细胞RNA测序（RNA-Seq）分析的技术方案。整体设计：为对比CEL-Seq2、CEL-Seq与Smart-Seq在C1平台上的性能，我们对小鼠树突状细胞与成纤维细胞进行了分析；我们使用秀丽隐杆线虫（C. elegans）总RNA对新型CEL-Seq2实验流程进行校准与优化。