文章基本信息

标题：Single-molecule denaturation mapping of DNA in nanofluidic channels
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作者：Walter Reisner ; Niels B. Larsen ; Asli Silahtaroglu 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2010
卷号：107
期号：30
页码：13294-13299
DOI：10.1073/pnas.1007081107
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：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.
关键词：DNA barcoding ; DNA denaturation ; DNA optical mapping ; nanochannel