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

Mater Sci Eng C Mater Biol Appl. 2017 May 1;74:10-20. doi: 10.1016/j.msec.2016.12.118. Epub 2017 Jan 10.

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 1.8-8.5×10-13m2 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; >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.

Keywords: Adsorption; DNA microextraction; Porous particles; Sorbent.

MeSH terms

  • Adsorption
  • DNA / blood
  • DNA / isolation & purification*
  • Genome, Human*
  • Humans
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Microscopy, Electron, Scanning
  • Particle Size
  • Polymers / chemistry*
  • Porosity
  • Silicon Dioxide / chemistry*

Substances

  • Polymers
  • Silicon Dioxide
  • DNA