School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, United States.
Anal Chem. 2011 Nov 1;83(21):8073-7. doi: 10.1021/ac202506j. Epub 2011 Oct 13.
We present a method for profiling the 5-methyl cytosine distribution on single DNA molecules. Our method combines soft-lithography and molecular elongation to form ordered arrays estimated to contain more than 250 000 individual DNA molecules immobilized on a solid substrate. The methylation state of the DNA is detected and mapped by binding of fluorescently labeled methyl-CpG binding domain peptides to the elongated dsDNA molecules and imaging of their distribution. The stretched molecules are fixed in their extended configuration by adsorption onto the substrate so analysis can be performed with high spatial resolution and signal averaging. We further prove this technique allows imaging of DNA molecules with different methylation states.
我们提出了一种在单链 DNA 分子上进行 5-甲基胞嘧啶分布分析的方法。我们的方法结合了软光刻和分子延伸技术，在固体基底上形成了估计含有 250000 个以上固定 DNA 分子的有序阵列。通过将荧光标记的甲基化 CpG 结合域肽与延伸的双链 DNA 分子结合，并对其分布进行成像，来检测和绘制 DNA 的甲基化状态。伸展的分子通过吸附到基底上而固定在其伸展的构象中，因此可以进行高空间分辨率和信号平均的分析。我们进一步证明了这种技术可以对具有不同甲基化状态的 DNA 分子进行成像。
