León-Paz-de-Rodríguez Gloria Elena, Rodríguez-León Ericka, Iñiguez-Palomares Ramón
Independent researcher, Hermosillo C.P. 83250, Sonora, México.
Physics Department, Universidad de Sonora, Hermosillo C.P. 83250, Sonora, México.
ACS Omega. 2024 Feb 12;9(8):9013-9026. doi: 10.1021/acsomega.3c07379. eCollection 2024 Feb 27.
This study presents a new procedure to condense DNA molecules and precipitate them onto a glass slide. The resulting DNA molecules undergo autonomous self-assembly, creating closed superstructures on the micrometer scale, which are called DNA hyperstructures. These structures can be observed using low-magnification (4×) light microscopy. Precisely controlling the alcohol/glacial acetic acid ratio and DNA concentration during precipitation enabled the regulation of structure compaction on the slide. The alcohol/glacial acetic acid ratio is inversely proportional to the DNA concentration to achieve optimal compaction on the slide. Confocal microscopy fluorescence analysis of DNA extracts stained with DAPI shows that nucleic acids self-assemble to form structures during precipitation on the slide. This methodology is relevant since it facilitates the precipitation and visualization of DNA, regardless of its origin or molecular weight. To confirm its versatility, results with DNA extracted from human peripheral blood, the Lambda virus, and plasmid pBR322 are presented. The study examined the morphological features of DNA hyperstructures in both healthy individuals and those diagnosed with different medical conditions or illnesses, revealing distinct patterns specific to each case. This innovative technology has potential for disease detection in peripheral blood samples, ranging from cancer and Alzheimer's disease to determining the gender of the gestational product at an early stage.
本研究提出了一种浓缩DNA分子并将其沉淀到载玻片上的新方法。所得的DNA分子进行自主自组装,在微米尺度上形成封闭的超结构,这些结构被称为DNA超结构。可以使用低倍(4×)光学显微镜观察这些结构。在沉淀过程中精确控制乙醇/冰醋酸比例和DNA浓度能够调节载玻片上结构的致密程度。乙醇/冰醋酸比例与DNA浓度成反比,以在载玻片上实现最佳致密程度。用DAPI染色的DNA提取物的共聚焦显微镜荧光分析表明,核酸在载玻片上沉淀过程中自组装形成结构。这种方法具有重要意义,因为它有助于DNA的沉淀和可视化,无论其来源或分子量如何。为了证实其通用性,展示了从人外周血、λ病毒和质粒pBR322中提取的DNA的结果。该研究检查了健康个体以及被诊断患有不同疾病或病症的个体中DNA超结构的形态特征,揭示了每种情况特有的不同模式。这项创新技术在从癌症和阿尔茨海默病到早期确定妊娠产物性别的外周血样本疾病检测中具有潜力。