Topography-assisted electromagnetic platform for blood-to-PCR in a droplet

Biosens Bioelectron. 2013 Dec 15:50:91-9. doi: 10.1016/j.bios.2013.06.011. Epub 2013 Jun 15.

Abstract

This paper presents an electromagnetically actuated platform for automated sample preparation and detection of nucleic acids. The proposed platform integrates nucleic acid extraction using silica-coated magnetic particles with real-time polymerase chain reaction (PCR) on a single cartridge. Extraction of genomic material was automated by manipulating magnetic particles in droplets using a series of planar coil electromagnets assisted by topographical features, enabling efficient fluidic processing over a variety of buffers and reagents. The functionality of the platform was demonstrated by performing nucleic acid extraction from whole blood, followed by real-time PCR detection of KRAS oncogene. Automated sample processing from whole blood to PCR-ready droplet was performed in 15 min. We took a modular approach of decoupling the modules of magnetic manipulation and optical detection from the device itself, enabling a low-complexity cartridge that operates in tandem with simple external instruments.

Keywords: Droplet manipulation; Micro-total-analysis system; Real-time PCR; Solid-phase DNA extraction.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / blood*
  • DNA / isolation & purification*
  • Electromagnetic Phenomena
  • Equipment Design
  • Humans
  • Magnets / chemistry
  • Microfluidic Analytical Techniques / instrumentation*
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins p21(ras)
  • Real-Time Polymerase Chain Reaction / instrumentation*
  • ras Proteins / genetics*

Substances

  • KRAS protein, human
  • Proto-Oncogene Proteins
  • DNA
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins