MANYETİK VE MANYETİK OLMAYAN SİLİKA BAZLI SABİT FAZLAR KULLANAN DNA MİKROEKSTRAKSİYON SİSTEMLERİNİN GELİŞTİRİLMESİ

Abstract

Monodisperse-porous silica microspheres and their magnetic form were synthesized by a newly developed” staged shape templated hydrolysis and condensation protocol”. The human genomic DNA isolation capacity from whole blood was analyzed with magnetic silica microspheres and bare silica microspheres as the sorbents in batch fashion and in a continous microextraction system, respectively. The pore size distribution and the specific surface area of microspheres were determined by nitrogen adsorption/desorption method. The microsphere size and the size distribution, the magnetic behaviour and the crystal structure were analyzed by scanning electron microscope, vibrating sample magnetometer and X-ray difractometer, respectively. First, the conditions were optimized for human genomic DNA isolation in batch fashion by using magnetic silica microspheres as the sorbent. Next, the bare silica microspheres were packed into a fused-silica capillary with 300 m i.d. and 40 mm in length. The microcolumn was evaluated for the isolation of genomic DNA from human whole blood via continous microextraction. The effects of lysate/adsorption buffer ratio and the sample volume on the DNA isolation yield and the amount of DNA isolated were determined in the microextraction system fabricated. Human genomic DNA samples acquired from whole blood by using both systems, were reproduced by polymerase chain reaction and then run on agarose gel electrophoresis. The results indicated that both the magnetic silica microspheres and the bare silica microspheres are promising alternative sobents for human genomic DNA isolation from human blood in batch fashion and continuous microextraction, respectively.