title: Human genomic DNA isolation from whole blood using a simple microfluidic system with silica- and polymer-based stationary phases.
abstract: Monodisperse-porous silica microspheres 5.1μm 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 1.8-8.5×10-13m2were 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; >2500ng of hgDNA was recovered with an isolation yield of about 50%, using an hgDNA feed concentration of 100ng/μL. Monodisperse-porous silica microspheres were also evaluated as a sorbent for genomic DNA isolation from human whole blood in the microfluidic system; 14ng of hgDNA was obtained from 10μL 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.
authors.affiliation: Hacettepe University, Bioengineering Division, Ankara, Turkey.
authors.forename: Gülçin
authors.initials: G
authors.lastname: Günal
authors.affiliation: Hacettepe University, Chemical Engineering Department, Ankara, Turkey.
authors.forename: Çiğdem
authors.initials: Ç
authors.lastname: Kip
authors.affiliation: Hacettepe University, Bioengineering Division, Ankara, Turkey.
authors.forename: Sevim Eda
authors.initials: SE
authors.lastname: Öğüt
authors.affiliation: Gazi University, Division of Medical Biology & Genetics, Ankara, Turkey.
authors.forename: Duygu Deniz
authors.initials: DD
authors.lastname: Usta
authors.affiliation: Hacettepe University, Division of Nanotechnology and Nanomedicine, Ankara, Turkey.
authors.forename: Erhan
authors.initials: E
authors.lastname: Şenlik
authors.affiliation: Hacettepe University, Bioengineering Division, Ankara, Turkey.
authors.forename: Güneş
authors.initials: G
authors.lastname: Kibar
authors.affiliation: Hacettepe University, Bioengineering Division, Ankara, Turkey; Hacettepe University, Division of Nanotechnology and Nanomedicine, Ankara, Turkey. Electronic address: atuncel@hacettepe.edu.tr.
authors.forename: Ali
authors.initials: A
authors.lastname: Tuncel
mesh_terms: D000327:Adsorption; D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D015894:Genome, Human; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D010316:Particle Size; D011108:Polymers / Q000737:chemistry*; D016062:Porosity; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D011108:Polymers; D012822:Silicon Dioxide; D004247:DNA
keywords: Adsorption; DNA microextraction; Porous particles; Sorbent