Methylation in DNA is a controlling factor in gene expression, embryonic development, and has been found to be important in infections and cancer. From a basic biology point of view, great heterogeneity has been found in methylation levels within tissues, so questions arises as to how and why. We show that methylated-DNA (m-DNA) can be distinguished from non-methylated (n-DNA) with nano-bowtie- and resonance- enhanced Raman spectra. By tuning the bowtie antenna to the resonance wavelength, both gains can be realized. Two additional Raman peaks in the 1200 – 1700 cm-1 band appear with methylation: one at 1239 cm-1 and the other at 1639 cm-1; a weak peak near 1000 cm-1 also appears with methylation. We also find that the two spectral features, although the latter with slight modification, can be used to distinguish the methylation state even when the DNA is denatured, as we show when we induce crystallization of the salts in the solution with increased excitation power, or allow it to happen naturally via solvent evaporation, and the DNA is trapped within the salt crystals. A comparison between liquid/solution to dried/denatured state m-DNA shows a general broadening of the larger lines and a transfer of spectral weight from the ~1470 cm-1 vibration to two higher energy lines. The applicability of the resonance-Raman in these spectra is shown by demonstrating that the Raman spectral characteristics hardly change as the Raman resonance in excitation wavelength is approached. Finally, we comment on real signal gain in this double-resonance system.
Ling Li, Shuang Fang Lim, Alexander Puretzky, Brandon J. N. Long, Robert Riehn, and Hans Hallen, "DNA methylation detection using UV nano bowtie antenna enhanced Raman spectroscopy," Proc. SPIE 10727, UV and Higher Energy Photonics: From Materials to Applications 2018, 107270B (Presented at SPIE Nanoscience + Engineering: August 19, 2018; Published: 7 September 2018); https://doi.org/10.1117/12.2321283.
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