DNA methylation profiling in nanochannels
DNA methylation profiling in nanochannels
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DOI：
10.1063/1.3613671
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发表时间：
2011-09-01
期刊：
影响因子：
3.2
通讯作者：
Riehn, Robert
中科院分区：
文献类型：
--
作者：
Lim, Shuang Fang;Karpusenko, Alena;Riehn, Robert
我们报告的分析5-甲基胞嘧啶分布在单个基因组大小的DNA分子在基因相关的决议。这种方法通过限制在尺寸约为250 x 200 nm的通道中来线性化和拉伸DNA分子（2）。使用荧光标记的甲基-CpG结合结构域蛋白（MBD）检测甲基化状态，具有高信号对比度和低背景。产生由甲基化和非甲基化区段组成的DNA条形码，其中短和长多联体两者均显示空间分辨的MBD结合。该技术的分辨率优于10 kbp，并已实现超过140 kbp的单分子读取长度。(C)2011年美国物理学会。[doi：10.1063/1.3613671]
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 x 200nm(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. (C) 2011 American Institute of Physics. [doi:10.1063/1.3613671]