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

Lotien Richard Huang, Jonas O. Tegenfeld, Jessica J. Kraeft, James C. Sturm, Robert H. Austin, Edward C. Cox

Research output: Contribution to journalArticle

188 Scopus citations

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 -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.

Original languageEnglish (US)
Pages (from-to)1048-1051
Number of pages4
JournalNature biotechnology
Volume20
Issue number10
DOIs
StatePublished - Oct 1 2002

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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