Human genomic DNA isolation from whole blood using a simple microfluidic system with silica- and polymer-based stationary phases


Gunal G., Kip C. , Ogut S. E. , Usta D. D. , ŞENLİK E. , Kibar G. , ...Daha Fazla

MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS, cilt.74, ss.10-20, 2017 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 74
  • Basım Tarihi: 2017
  • Doi Numarası: 10.1016/j.msec.2016.12.118
  • Dergi Adı: MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS
  • Sayfa Sayıları: ss.10-20

Özet

Monodisperse-porous silica microspheres 5.1 mu m in size with a bimodal pore-size distribution (including both mesoporous and macroporous compartments) were obtained using a newly developed staged-shape templated hydrolysis and condensation protocol. Synthesized silica microspheres and monodisperse-porous polymer based microspheres with different functionalities, synthesized by staged-shape template polymerization, were comparatively tested as sorbents for human genomic DNA (hgDNA) isolation in a microfluidic system. Microcolumns with a permeability range of 1.8-8.5 x 10(-13) m(2) were fabricated by the slurry-packing of silica or polymer-based microspheres. The monodisperse-porous silica microspheres showed the best performance in hgDNA isolation in an aqueous buffer medium; >2500 ng of hgDNA was recovered with an isolation yield of about 50%, using an hgDNA feed concentration of 100 ng/mu L. Monodisperse-porous silica microspheres were also evaluated as a sorbent for genomic DNA isolation from human whole blood in the microfluidic system; 14 ng of hgDNA was obtained from 10 mu L. of whole blood lysate with an isolation yield of 64%. Based on these results, we conclude that monodisperse-porous silica microspheres with a bimodal pore size distribution are a promising sorbent for the isolation of hgDNA in larger amounts and with higher yields compared to the sorbents previously tried in similar microfluidic systems. (C) 2017 Elsevier B.V. All rights reserved.