Centre for Biophotonics, Lancaster Environment Centre, Lancaster University, Bailrigg, Lancaster LA1 4YQ, UK.
J Biophotonics. 2011 May;4(5):345-54. doi: 10.1002/jbio.201000085. Epub 2010 Oct 29.
Methylation status plays important roles in the regulation of gene expression and significantly influences the dynamics, bending and flexibility of DNA. The aim of this study was to determine whether attenuated total reflection Fourier-transform infrared (ATR-FTIR) or Raman spectroscopy with subsequent multivariate analysis could determine methylation patterning in oligonucleotides variously containing 5-methylcytosine, cytosine and guanine bases. Applied to Low-E reflective glass slides, 10 independent spectral acquisitions were acquired per oligonucleotide sample. Resultant spectra were baseline-corrected and vector normalised over the 1750 cm(-1) -760 cm(-1) (for ATR-FTIR spectroscopy) or the 1750 cm(-1) -600 cm(-1) (for Raman spectroscopy) regions. Data were then analysed using principal component analysis (PCA) coupled with linear discriminant analysis (LDA). Exploiting this approach, biomolecular signatures enabling sensitive and specific discrimination of methylation patterning were derived. For DNA sequence and methylation analysis, this approach has the potential to be an important tool, especially when material is scarce.
甲基化状态在基因表达调控中起着重要作用，显著影响 DNA 的动态、弯曲和柔韧性。本研究旨在确定衰减全反射傅里叶变换红外（ATR-FTIR）或拉曼光谱分析与多元分析相结合是否可以确定各种含有 5-甲基胞嘧啶、胞嘧啶和鸟嘌呤碱基的寡核苷酸的甲基化模式。将 10 个独立的光谱采集应用于低辐射反射玻璃载玻片上，每个寡核苷酸样本采集 10 次。所得光谱在 1750 cm(-1) -760 cm(-1)（ATR-FTIR 光谱学）或 1750 cm(-1) -600 cm(-1)（拉曼光谱学）区域进行基线校正和向量归一化。然后使用主成分分析（PCA）结合线性判别分析（LDA）对数据进行分析。利用这种方法，可以得出能够灵敏和特异性区分甲基化模式的生物分子特征。对于 DNA 序列和甲基化分析，这种方法有可能成为一种重要的工具，特别是在材料稀缺的情况下。
