On-line cell lysis and DNA extraction on a microfluidic biochip fabricated by microelectromechanical system technology

Electrophoresis. 2008 May;29(9):1844-51. doi: 10.1002/elps.200700551.

Abstract

Integrating cell lysis and DNA purification process into a micrototal analytical system (microTAS) is one critical step for the analysis of nucleic acids. On-chip cell lysis based on a chemical method is realized by sufficient blend of blood sample and the lyzing reagent. In this paper two mixing models, T-type mixing model and sandwich-type mixing model, are proposed and simulation of those models is conducted. Result of simulation shows that the sandwich-type mixing model with coiled channel performs best and this model is further used to construct the microfluidic biochip for on-line cell lysis and DNA extraction. The result of simulation is further verified by experiments. It asserts that more than 80% mixing of blood sample and lyzing reagent which guarantees that completed cell lysis can be achieved near the inlet location when the cell/buffer velocity ratio is less than 1:5. After cell lysis, DNA extraction by means of a solid-phase method is implemented by using porous silicon matrix which is integrated in the biochip. During continuous flow process in the microchip, rapid cell lysis and PCR-amplifiable genomic DNA purification can be achieved within 20 min. The potential of this microfluidic biochip is illustrated by pretreating a whole blood sample, which shows the possibility of integration of sample preparation, PCR, and separation on a single device to work as portable point-of-care medical diagnostic system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzothiazoles
  • Blood Cells / chemistry*
  • Buffers
  • DNA / blood
  • DNA / isolation & purification*
  • Diamines
  • Electrochemistry
  • Equipment Design
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods
  • Fluorescent Dyes
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Models, Chemical
  • Organic Chemicals
  • Polymerase Chain Reaction
  • Porosity
  • Quinolines
  • Rats
  • Silicon

Substances

  • Benzothiazoles
  • Buffers
  • Diamines
  • Fluorescent Dyes
  • Organic Chemicals
  • Quinolines
  • SYBR Green I
  • DNA
  • Silicon