期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 107, 期 30, 页码 13294-13299出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1007081107
关键词
DNA barcoding; DNA denaturation; DNA optical mapping; nanochannel
资金
- Danish Research Council for Technology and Production Sciences [274-05-0375]
- Human Frontier Science Program [RGY78/2007]
- Swedish Research Council [2002-5972, 2007-584]
- Wilhelm Johannsen Centre for Functional Genome Research
- European Community [HEALTH-F4-2008-201418]
Here we explore the potential power of denaturation mapping as a single-molecule technique. By partially denaturing YOYO (R)-1-labeled DNA in nanofluidic channels with a combination of formamide and local heating, we obtain a sequence-dependent barcode corresponding to a series of local dips and peaks in the intensity trace along the extended molecule. We demonstrate that this structure arises from the physics of local denaturation: statistical mechanical calculations of sequence-dependent melting probability can predict the barcode to be observed experimentally for a given sequence. Consequently, the technique is sensitive to sequence variation without requiring enzymatic labeling or a restriction step. This technique may serve as the basis for a new mapping technology ideally suited for investigating the long-range structure of entire genomes extracted from single cells.
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