A simultaneous space sampling method for DNA fraction collection using a comb structure in microfluidic devices

Electrophoresis. 2011 Nov;32(23):3392-8. doi: 10.1002/elps.201100362. Epub 2011 Nov 10.

Abstract

Fraction collection of selected components from a complex mixture plays a critical role in biomedical research, environmental analysis, and biotechnology. Here, we introduce a novel electrophoretic chip device based on a signal processing theorem that allows simultaneous space sampling for fractionation of ssDNA target fragments. Ten parallel extraction channels, which covered 1.5-mm-long sampling ranges, were used to facilitate the capturing of fast-moving fragments. Furthermore, the space sampling extraction made it possible to acquire pure collection, even from partly overlapping fragments that had been insufficiently separated after a short electrophoretic run. Fragments of 180, 181, and 182 bases were simultaneously collected, and then the recovered DNA was PCR amplified and assessed by CE analysis. The 181-base target was shown to be isolated in a 70-mm-long separation length within 10 min, in contrast to the >50 min required for the 300-mm-long separation channel in our previous study. This method provides effective combination of time and space, which is a breakthrough in the traditional concept of fraction collection on a chip.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / analysis
  • DNA / chemistry
  • DNA / isolation & purification*
  • Electrophoresis, Capillary / methods*
  • Equipment Design
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Polymerase Chain Reaction
  • Signal Processing, Computer-Assisted
  • Spectrometry, Fluorescence
  • Time Factors

Substances

  • DNA