Abstract
Bisulfite genomic sequencing provides a single-molecule view of cytosine methylation states. After deamination, each cloned molecule contains a record of methylation within its sequence. The full power of this technique is harnessed by treating nuclei with an exogenous DNMT prior to DNA extraction. This exogenous methylation marks regions of accessibility and footprints nucleosomes, as well as other DNA-binding proteins. Thus, each cloned molecule records not only the endogenous methylation present (at CG sites, in mammals), but also the exogenous (GC, when using the Chlorella virus protein M.CviPI). We term this technique MAPit, methylation accessibility protocol for individual templates.
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Acknowledgments
We are grateful to Amber Delmas for advice on troubleshooting BGS. This work was supported by the National Institutes of Health (CA95525 to MPK) as well as the Department of Defense, Breast Cancer Research Program (BC062914, BC087311, and BC097648 to MPK).
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Darst, R.P. et al. (2012). Simultaneous Single-Molecule Detection of Endogenous C-5 DNA Methylation and Chromatin Accessibility Using MAPit. In: Morse, R. (eds) Chromatin Remodeling. Methods in Molecular Biology, vol 833. Humana Press. https://doi.org/10.1007/978-1-61779-477-3_9
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DOI: https://doi.org/10.1007/978-1-61779-477-3_9
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