Department of Physics, North Carolina State University, Raleigh, North Carolina 27695, USA.
Biomicrofluidics. 2011 Sep;5(3):34106-341068. doi: 10.1063/1.3613671. Epub 2011 Jul 25.
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.
我们报告了在基因相关分辨率下对单个基因组大小的 DNA 分子内 5-甲基胞嘧啶分布的分析。该方法通过限制在尺寸约为 250×200nm(2)的通道中，使 DNA 分子线性化和拉伸。通过使用荧光标记的甲基-CpG 结合域蛋白 (MBD) 检测甲基化状态，具有高信号对比度和低背景。生成由甲基化和非甲基化片段组成的 DNA 条码，短和长串联物都表现出空间分辨的 MBD 结合。该技术的分辨率优于 10 kbp，并且已经实现了超过 140 kbp 的单分子读长。
