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Simultaneous single-molecule detection of endogenous C-5 DNA methylation and chromatin accessibility using MAPit.

Authors :
Darst RP
Pardo CE
Pondugula S
Gangaraju VK
Nabilsi NH
Bartholomew B
Kladde MP
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2012; Vol. 833, pp. 125-41.
Publication Year :
2012

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.

Details

Language :
English
ISSN :
1940-6029
Volume :
833
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
22183592
Full Text :
https://doi.org/10.1007/978-1-61779-477-3_9