New methods for DNA oligomer-binding and detection in the DNA microarray technology

摘要

Widely used molecular-chips of today are expression type DNA-microarryas. 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 polypirrol modified gold electrode. After immobilisation of biotin labeled DNA on electrode the detection was performed through streptavidine-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 fluorescense microscope. We investigated the influence of the polimerisation time to thickness and quality of the polimer 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.