期刊
NATURE PROTOCOLS
卷 10, 期 5, 页码 645-659出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2015.039
关键词
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资金
- National Science Foundation of China [31322037, 31271543]
- National Basic Research Program of China [2012CB966704, 2011CB966303]
- Center for Molecular and Translational Medicine
The heterogeneity of DNA methylation within a population of cells necessitates DNA methylome profiling at single-cell resolution. Recently, we developed a single-cell reduced-representation bisulfite sequencing (scRRBS) technique in which we modified the original RRBS method by integrating all the experimental steps before PCR amplification into a single-tube reaction. These modifications enable scRRBS to provide digitized methylation information on similar to 1 million CpG sites within an individual diploid mouse or human cell at single-base resolution. Compared with the single-cell bisulfite sequencing (scBS) technique, scRRBS covers fewer CpG sites, but it provides better coverage for CpG islands (CGIs), which are likely to be the most informative elements for DNA methylation. The entire procedure takes similar to 3 weeks, and it requires strong molecular biology skills.
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