DNA methylation profiling in nanochannels
| Autor: | Shuang Fang Lim, Tyra A. Lamar, Joseph A. Hook, Alena Karpusenko, John J. Sakon, Robert Riehn |
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| Rok vydání: | 2011 |
| Předmět: |
Fluid Flow and Transfer Processes
Concatemer Molecular biophysics Biomedical Engineering Methylation Biology Condensed Matter Physics Molecular biology Dna methylation profiling chemistry.chemical_compound Colloid and Surface Chemistry chemistry Dna barcodes Biophysics General Materials Science Cytosine DNA Regular Articles Binding domain |
| Zdroj: | Biomicrofluidics. 5:034106 |
| ISSN: | 1932-1058 |
| DOI: | 10.1063/1.3613671 |
| Popis: | We report the profiling of the 5-methyl cytosine distribution within single genomic-sized DNA molecules at a gene-relevant resolution. This method linearizes and stretches DNA molecules by confinement to channels with a dimension of about 250×200 nm(2). The methylation state is detected using fluorescently labeled methyl-CpG binding domain proteins (MBD), with high signal contrast and low background. DNA barcodes consisting of methylated and non-methylated segments are generated, with both short and long concatemers demonstrating spatially resolved MBD binding. The resolution of the technique is better than 10 kbp, and single-molecule read-lengths exceeding 140 kbp have been achieved. |
| Databáze: | OpenAIRE |
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