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

摘要

Monodisperse-porous silica microspheres 5.1 μm 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 1.8-8.5×10-13m2 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/μL. 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 μL 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.