Solid phase extraction of DNA from biological samples in a post-based, high surface area poly(methyl methacrylate) (PMMA) microdevice

Lab Chip. 2011 May 7;11(9):1603-11. doi: 10.1039/c0lc00597e. Epub 2011 Mar 4.

Abstract

This work describes the performance of poly(methyl methacrylate) (PMMA) microfluidic DNA purification devices with embedded microfabricated posts, functionalized with chitosan. PMMA is attractive as a substrate for creating high surface area (SA) posts for DNA capture because X-ray lithography can be exploited for extremely reproducible fabrication of high SA structures. However, this advantage is offset by the delicate nature of the posts when attempting bonding to create a closed system, and by the challenge of functionalizing the PMMA surface with a group that invokes DNA binding. Methods are described for covalent functionalization of the post surfaces with chitosan that binds DNA in a pH-dependent manner, as well as for bonding methods that avoid damaging the underlying post structure. A number of geometric posts designs are explored, with the goal of identifying post structures that provide the requisite surface area without a concurrent rise in fluidic resistance that promotes device failure. Initial proof-of-principle is shown by recovery of prepurified human genomic DNA (hgDNA), with real-world utility illustrated by purifying hgDNA from whole blood and demonstrating it to be PCR-amplifiable.

MeSH terms

  • Blood Chemical Analysis / instrumentation
  • Blood Chemical Analysis / methods
  • Chitosan / chemistry
  • DNA / blood
  • DNA / isolation & purification*
  • Equipment Design
  • Humans
  • Hydrogen-Ion Concentration
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Polymethyl Methacrylate / chemistry*
  • Solid Phase Extraction / methods*
  • Surface Properties

Substances

  • DNA
  • Polymethyl Methacrylate
  • Chitosan