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Abstract
The genome-wide DNA methylation profile, or DNA methylome, is a critical component of the overall epigenomic landscape that modulates gene activities and cell fate. Single-cell DNA methylomic studies offer unprecedented resolution for detecting and profiling cell subsets based on methylomic features. However, existing single-cell methylomic technologies are based on use of tubes or well plates and these platforms are not easily scalable for handling a large number of single cells. Here we demonstrate a droplet-based microfluidic technology, Drop-BS, to construct single-cell bisulfite sequencing libraries for DNA methylome profiling. Drop-BS takes advantage of the ultrahigh throughput offered by droplet microfluidics to prepare bisulfite sequencing libraries of up to 10,000 single cells within 2 days. We apply the technology to profile mixed cell lines, mouse and human brain tissues to reveal cell type heterogeneity. Drop-BS offers a promising solution for single-cell methylomic studies requiring examination of a large cell population.
Single-cell DNA methylomic studies offer high resolution to differentiate cell subsets based on their epigenomic features. Here, the authors demonstrate Drop-BS, a droplet-based single-cell bisulfite sequencing library preparation method, for DNA methylome profiling.
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1 Virginia Tech, Department of Chemical Engineering, Blacksburg, USA (GRID:grid.438526.e) (ISNI:0000 0001 0694 4940)
2 Virginia Tech, Department of Biomedical Engineering and Mechanics, Blacksburg, USA (GRID:grid.438526.e) (ISNI:0000 0001 0694 4940); Icahn School of Medicine at Mount Sinai, Department of Genetics and Genomic Sciences, New York, USA (GRID:grid.59734.3c) (ISNI:0000 0001 0670 2351)
3 Virginia Tech, Department of Biomedical Engineering and Mechanics, Blacksburg, USA (GRID:grid.438526.e) (ISNI:0000 0001 0694 4940)
4 University of the Basque Country UPV/EHU, CIBERSAM, Biocruces Health Research Institute, Department of Pharmacology, Leioa, Spain (GRID:grid.11480.3c) (ISNI:0000000121671098)
5 Virginia Commonwealth University School of Medicine, Department of Physiology and Biophysics, Richmond, USA (GRID:grid.224260.0) (ISNI:0000 0004 0458 8737)