1Beijing Advanced Innovation Center for Genomics (ICG), Biomedical Pioneering Innovation Center (BIOPIC), School of Life Sciences, College of Engineering, and Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, 100871 China.
2School of Life Sciences, and Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, 100084 China.
Commun Biol. 2019 Apr 29;2:147. doi: 10.1038/s42003-019-0401-y. eCollection 2019.
Single-cell whole-genome sequencing (scWGS) is mainly used to probe intercellular genomic variations, focusing on the copy number variations or alterations and the single-nucleotide variations (SNVs) occurring within single cells. Single-cell whole-genome amplification (scWGA) needs to be applied before scWGS but is challenging due to the low copy number of DNA. Besides, many genomic variations are rare within a population of cells, so the throughput of currently available scWGA methods is far from satisfactory. Here, we integrate a one-step micro-capillary array (MiCA)-based centrifugal droplet generation technique with emulsion multiple displacement amplification (eMDA) and demonstrate a high-throughput scWGA method, MiCA-eMDA. MiCA-eMDA increases the single-run throughput of scWGA to a few dozen, and enables the assessment of copy number variations and alterations at 50-kb resolution. Downstream target enrichment further enables the detection of SNVs with 20% allele drop-out.
单细胞全基因组测序 (scWGS) 主要用于探测细胞间基因组变异，侧重于单个细胞内发生的拷贝数变异或改变以及单核苷酸变异 (SNVs)。scWGS 之前需要进行单细胞全基因组扩增 (scWGA)，但由于 DNA 拷贝数低，这具有挑战性。此外，许多基因组变异在细胞群体中是罕见的，因此目前可用的 scWGA 方法的通量远远不能令人满意。在这里，我们将一步式微毛细管阵列 (MiCA) 为基础的离心液滴生成技术与乳液多重置换扩增 (eMDA) 相结合，展示了一种高通量的 scWGA 方法，MiCA-eMDA。MiCA-eMDA 将 scWGA 的单轮通量提高到几十个，并且能够以 50-kb 的分辨率评估拷贝数变异和改变。下游目标富集进一步使检测具有 20%等位基因缺失的 SNVs 成为可能。
