DNA微阵列技术中DNA寡聚物结合与检测的新方法

A. Győrffy
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引用次数: 0

摘要

目前广泛使用的分子芯片是表达型dna微阵列。由于成本高、需要大量的实验室设备以及结果不明确,这些芯片在临床实践中并不常用。因此,对简单的大规模生产方法的需求不断增长。在我们的工作中,我们开发了一种新的DNA固定方法,用于DNA在聚吡咯修饰金电极上的结合。将生物素标记的DNA固定在电极上后,通过链霉亲和素-德克萨斯红染料结合进行检测。未标记的DNA探针用他的苯并胺染料检测。在检测中,我们使用共聚焦荧光显微镜对DNA进行可视化。我们研究了聚合时间对聚合物膜厚度和质量的影响,以及用于检测和固定的DNA数量的影响。在我们的构想中,我们的目标是构建一种临床适用的芯片,用于鉴定PCR扩增的dna,用于未来遗传病的已知突变。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
New methods of DNA oligomer - binding and detection in the DNA microarray technology
Widely used molecular-chips of today are expression type DNA-microarrays. These chips are not commonly used in clinical practice due to the high cost, the need for extensive laboratory devices and due to not well clarified results. Therefore the need for simple mass-production methods is constantly growing. In our work we have developed a new DNA immobilisation method for DNA binding on polypyrrole modified gold electrode. After immobilisation of biotin labeled DNA on electrode the detection was performed through streptavidin-texas red dye binding. The unlabeled DNA probes were detected using bis benzimide dye. For the visualisation of the DNA in detection we used confocal fluorescence microscope. We investigated the influence of the polymerisation time to thickness and quality of the polymer film and the influence of the quantity of DNA for detection and immobilisation. In our conception we targeted to build a clinically applicable chip, to be used for the identification of PCR amplified DNAs for known mutations of inherited diseases in the future.
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