A DNA prism for high-speed continuous fractionation of large DNA molecules

Nat Biotechnol. 2002 Oct;20(10):1048-51. doi: 10.1038/nbt733. Epub 2002 Sep 3.

Abstract

The analysis and fractionation of large DNA molecules plays a key role in many genome projects. The standard method, pulsed-field gel electrophoresis (PFGE), is slow, with running times ranging from 10 hours to more than 200 hours. In this report, we describe a thumbnail-sized device that sorts large DNA fragments (61-209 kilobases (kb)) in 15 seconds, with a resolution of approximately 13%. An array of micron-scale posts serves as the sieving matrix, and integrated microfluidic channels spatially shape the electric fields over the matrix. Asymmetric pulsed fields are applied for continuous-flow operation, which sorts DNA molecules in different directions according to their molecular masses, much as a prism deflects light of different wavelengths at different angles. We demonstrate the robustness of the device by using it to separate large DNA inserts prepared from bacterial artificial chromosomes, a widely used DNA source for most genomics projects.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Technical Report

MeSH terms

  • Chemical Fractionation / instrumentation*
  • Chemical Fractionation / methods*
  • Chromosomes, Artificial, Bacterial
  • Chromosomes, Artificial, P1 Bacteriophage
  • DNA, Bacterial / analysis*
  • Electromagnetic Fields
  • Electrophoresis / methods*
  • Nanotechnology
  • Oligonucleotide Array Sequence Analysis / instrumentation*
  • Oligonucleotide Array Sequence Analysis / methods*

Substances

  • DNA, Bacterial