scSPLAT, a scalable plate-based protocol for single cell WGBS library preparation

Sci Rep. 2022 Apr 6;12(1):5772. doi: 10.1038/s41598-022-09798-2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Methylation
  • Gene Library
  • High-Throughput Nucleotide Sequencing* / methods
  • Oligonucleotides
  • Sequence Analysis, DNA / methods
  • Sulfites*

Substances

  • Oligonucleotides
  • Sulfites