MANYETİK VE MANYETİK OLMAYAN TiO2 BAZLI SABİT FAZLAR İLE GENOMİK DNA İZOLASYONU

Abstract

The aim of this thesis is the synthesis of monodisperse porous magnetic and non-magnetic titania (titanium dioxide) microspheres with high genomic DNA isolation ability and investigation of genomic DNA isolation performance in batch fashion. At the first stage, monodisperse porous non-magnetic and magnetic titania microspheres were synthesized using the sol-gel method. For synthesis of non-magnetic titania particles, poly (HPMA-Cl-co-EDMA) particles were derivatized by sodium bisulfite and subsequently were utilized as a template in the presence of titanium chloride precursor. Magnetic titania microspheres were synthesized by primarily derivatization using ethylene diamine and subsequently were magnetized by the binary precipitation method using iron salts. The magnetic poly (HPMA-Cl-co-EDMA) microspheres were used as a template and titanium chloride as a precursor. In the second stage, genomic DNA isolation performances of monodisperse porous magnetic and non-magnetic titania microspheres were investigated using calf thymus and human genomic DNA. The effect of type of desorption medium, pH of adsorption medium, initial DNA concentration and amount of sorbent on DNA isolation were performed in batch fashion. By considering the above optimized parameters, the effect of sorbent amount on DNA isolation from whole blood was investigated using non-magnetic titania microspheres. Isolated human genomic DNA was characterized using polymerase chain reaction (PCR) and agarose gel electrophoresis methods. The obtained results showed that monodisperse porous magnetic and non-magnetic titania microspheres exhibited satisfactory genomic DNA isolation performance compared to the conventionally available sorbents and could be offered as an alternative solid phase for solid phase DNA isolation in batch fashion.