Ruan Qingyu, Ruan Weidong, Lin Xiaoye, Wang Yang, Zou Fenxiang, Zhou Leiji, Zhu Zhi, Yang Chaoyong
Collaborative Innovation Center of Chemistry for Energy Materials, MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemical Engineering, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, P.R. China.
Institute of Molecular Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China.
Sci Adv. 2020 Dec 9;6(50). doi: 10.1126/sciadv.abd6454. Print 2020 Dec.
Single-cell whole-genome sequencing (WGS) is critical for characterizing dynamic intercellular changes in DNA. Current sample preparation technologies for single-cell WGS are complex, expensive, and suffer from high amplification bias and errors. Here, we describe Digital-WGS, a sample preparation platform that streamlines high-performance single-cell WGS with automatic processing based on digital microfluidics. Using the method, we provide high single-cell capture efficiency for any amount and types of cells by a wetted hydrodynamic structure. The digital control of droplets in a closed hydrophobic interface enables the complete removal of exogenous DNA, sufficient cell lysis, and lossless amplicon recovery, achieving the low coefficient of variation and high coverage at multiple scales. The single-cell genomic variations profiling performs the excellent detection of copy number variants with the smallest bin of 150 kb and single-nucleotide variants with allele dropout rate of 5.2%, holding great promise for broader applications of single-cell genomics.
单细胞全基因组测序（WGS）对于表征DNA中的动态细胞间变化至关重要。当前用于单细胞WGS的样本制备技术复杂、昂贵，且存在高扩增偏差和错误。在此，我们描述了数字WGS，这是一种样本制备平台，基于数字微流控通过自动处理简化了高性能单细胞WGS。使用该方法，我们通过湿润的流体动力学结构为任何数量和类型的细胞提供了高单细胞捕获效率。在封闭的疏水界面中对液滴进行数字控制能够完全去除外源DNA，实现充分的细胞裂解和无损扩增子回收，在多个尺度上实现低变异系数和高覆盖率。单细胞基因组变异分析能够出色地检测最小区间为150 kb的拷贝数变异以及等位基因缺失率为5.2%的单核苷酸变异，为单细胞基因组学的更广泛应用带来了巨大希望。
