Science for Life Laboratory, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
Sci Rep. 2022 Apr 6;12(1):5772. doi: 10.1038/s41598-022-09798-2.
DNA methylation is a central epigenetic mark that has diverse roles in gene regulation, development, and maintenance of genome integrity. 5 methyl cytosine (5mC) can be interrogated at base resolution in single cells by using bisulfite sequencing (scWGBS). Several different scWGBS strategies have been described in recent years to study DNA methylation in single cells. However, there remain limitations with respect to cost-efficiency and yield. Herein, we present a new development in the field of scWGBS library preparation; single cell Splinted Ligation Adapter Tagging (scSPLAT). scSPLAT employs a pooling strategy to facilitate sample preparation at a higher scale and throughput than previously possible. We demonstrate the accuracy and robustness of the method by generating data from 225 single K562 cells and from 309 single liver nuclei and compare scSPLAT against other scWGBS methods.
DNA 甲基化是一种重要的表观遗传标记，在基因调控、发育和基因组完整性维持中具有多种作用。5-甲基胞嘧啶（5mC）可以通过亚硫酸氢盐测序（scWGBS）在单细胞中以碱基分辨率进行检测。近年来，已经描述了几种不同的 scWGBS 策略来研究单细胞中的 DNA 甲基化。然而，在成本效益和产量方面仍然存在局限性。在这里，我们提出了 scWGBS 文库制备领域的一项新进展；单细胞 Splinted Ligation Adapter Tagging（scSPLAT）。scSPLAT 采用一种池化策略，以比以前更高的规模和通量促进样品制备。我们通过从 225 个单个 K562 细胞和 309 个单个肝细胞核生成数据来证明该方法的准确性和稳健性，并将 scSPLAT 与其他 scWGBS 方法进行比较。
