Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland.
Department of Industrial Products and Packaging Quality, Poznań University of Economics and Business, Poznań, Poland.
PLoS One. 2024 Jun 14;19(6):e0305590. doi: 10.1371/journal.pone.0305590. eCollection 2024.
The analysis of nucleic acids is one of the fundamental parts of modern molecular biology and molecular diagnostics. The information collected predominantly depends on the condition of the genetic material. All potential damage induced by oxidative stress may affect the final results of the analysis of genetic material obtained using commonly used techniques such as polymerase chain reaction or sequencing. The aim of this work was to evaluate the effects of high temperature and pH on DNA structure in the context of the occurrence of oxidative damage, using square-wave voltammetry and two independent research protocols. We resulted in visible oxidation damage registered in acidic conditions after the thermal denaturation process (pH 4.7) with changes in the intensity of guanine and adenine signals. However, using phosphate buffer (pH 7.0) for DNA denaturation negatively affected the DNA structure, but without any oxidized derivatives present. This leads to the conclusion that oxidation occurring in the DNA melting process results in the formation of various derivatives of nucleobases, both electrochemically active and inactive. These derivatives may distort the results of molecular tests due to the possibility of forming complementary bonds with various nucleobases. For example, 8-oxoguanine can form pairs with both cytosine and adenine.
核酸分析是现代分子生物学和分子诊断学的基础部分之一。收集的信息主要取决于遗传物质的状况。氧化应激引起的所有潜在损伤都可能影响使用聚合酶链反应或测序等常用技术获得的遗传物质分析的最终结果。本工作的目的是使用方波伏安法和两种独立的研究方案，评估高温和 pH 值对氧化损伤发生时 DNA 结构的影响。我们在热变性过程（pH 4.7）后酸性条件下观察到可见的氧化损伤，鸟嘌呤和腺嘌呤信号的强度发生变化。然而，使用磷酸盐缓冲液（pH 7.0）进行 DNA 变性会对 DNA 结构产生负面影响，但不存在任何氧化衍生物。这得出结论，即在 DNA 熔融过程中发生的氧化会导致形成各种碱基的衍生物，既有电化学活性的也有无活性的。这些衍生物可能会由于与各种碱基形成互补键的可能性而扭曲分子测试的结果。例如，8-氧鸟嘌呤可以与胞嘧啶和腺嘌呤形成碱基对。
