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Sci Rep. 2013;3:1389. doi: 10.1038/srep01389.

Detection and quantification of methylation in DNA using solid-state nanopores.

Author information

1
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA.

Abstract

Epigenetic modifications in eukaryotic genomes occur primarily in the form of 5-methylcytosine (5 mC). These modifications are heavily involved in transcriptional repression, gene regulation, development and the progression of diseases including cancer. We report a new single-molecule assay for the detection of DNA methylation using solid-state nanopores. Methylation is detected by selectively labeling methylation sites with MBD1 (MBD-1x) proteins, the complex inducing a 3 fold increase in ionic blockage current relative to unmethylated DNA. Furthermore, the discrimination of methylated and unmethylated DNA is demonstrated in the presence of only a single bound protein, thereby giving a resolution of a single methylated CpG dinucleotide. The extent of methylation of a target molecule could also be coarsely quantified using this novel approach. This nanopore-based methylation sensitive assay circumvents the need for bisulfite conversion, fluorescent labeling, and PCR and could therefore prove very useful in studying the role of epigenetics in human disease.

PMID:
23474808
PMCID:
PMC3593219
DOI:
10.1038/srep01389
[Indexed for MEDLINE]
Free PMC Article
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