1. Dual detection of chromatin accessibility and DNA methylation using ATAC-Me.
- Author
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Guerin LN, Barnett KR, and Hodges E
- Subjects
- B-Lymphocytes cytology, B-Lymphocytes metabolism, Cell Differentiation, Cell Line, Cell Line, Tumor, Chromatin chemistry, CpG Islands, DNA genetics, DNA Transposable Elements, Gene Library, Human Embryonic Stem Cells cytology, Human Embryonic Stem Cells metabolism, Humans, Software, Sulfites chemistry, THP-1 Cells, Transcription, Genetic, Transposases genetics, Transposases metabolism, Biological Assay, Chromatin metabolism, Computational Biology methods, DNA metabolism, DNA Methylation, Epigenesis, Genetic
- Abstract
The epigenome is multidimensional, with individual molecular components operating on different levels to control transcriptional output. Techniques that combine measurements of these features can reveal their precise correspondence in genomic space, or temporal connectivity, to better understand how they jointly regulate genes. ATAC-Me is an integrated method to probe DNA methylation and chromatin accessibility from a single DNA fragment library. Intact nuclei undergo Tn5 transposition to isolate DNA fragments within nucleosome-free regions. Isolated fragments are exposed to sodium bisulfite before library amplification and sequencing. A typical ATAC-Me experiment detects ~60,000-75,000 peak regions (P < 0.05), covering ~3-4 million CpG sites with at least 5× coverage. These sites display a range of methylation values depending on the cellular and genomic context. The approach is well suited for time course studies that aim to capture chromatin and DNA methylation dynamics in tandem during cellular differentiation. The protocol is completed in 2 d with standard molecular biology equipment and expertise. Analysis of resulting data uses publicly available software requiring basic bioinformatics skills to interpret results., (© 2021. Springer Nature Limited.)
- Published
- 2021
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