Single-cell whole-genome sequencing (scWGS) plays a critical role in revealing intercellular genome variations. However, the existing scWGS technologies suffer from tedious single-cell isolation and nonuniform amplification. Here, we present an Integrated single-tube operation for rapid and uniform single-cell whole-genome sequencing (Iso-seq) to overcome these limitations. The specially designed dual-well structure in the cap is based on size exclusion and locally quasi-hydrodynamic principles, which ensure that one cell is trapped in one cap, while the capillary-based centrifugal-driven droplet generator facilitates uniform droplet generation in the tube. Using Iso-seq, single cells can be isolated by the dual-well structure in an eight-cap strip within 3 min and the corresponding whole-genome of single cells can be readily compartmentalized in parallel droplets. The prepared single-cell samples show a high mapping ratio, broad coverage of the genome, and low coefficient of variation, demonstrating the good performance of Iso-seq in scWGS. With the advantages of rapid single-cell isolation, convenient droplet generation, broad coverage of the genome, and excellent sequencing uniformity, Iso-seq is promising for a wide variety of biological applications.

单细胞全基因组测序 (scWGS) 在揭示细胞间基因组变异方面起着关键作用。然而，现有的 scWGS 技术存在单细胞分离繁琐和扩增不均匀的问题。在这里，我们提出了一个集成的单管o用于快速和统一的单细胞全基因组测序（Iso-seq）的操作以克服这些限制。帽内特殊设计的双孔结构基于尺寸排阻和局部准流体力学原理，确保一个细胞被困在一个帽内，而基于毛细管的离心驱动液滴发生器有助于在管内均匀产生液滴. 使用 Iso-seq，单细胞可以在 3 分钟内通过八帽带中的双孔结构分离，并且单细胞的相应全基因组可以很容易地划分为平行液滴。制备的单细胞样本显示出高映射率、广泛的基因组覆盖度和低变异系数，证明了 Iso-seq 在 scWGS 中的良好性能。凭借快速单细胞分离的优势，