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- Title
A DNA prism for high-speed continuous fractionation of large DNA molecules.
- Authors
Huang, Lotien Richard; Tegenfeldt, Jonas O.; Kraeft, Jessica J.; Sturm, James C.; Austin, Robert H.; Cox, Edward C.
- 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.
- Subjects
DNA; PRISMS
- Publication
Nature Biotechnology, 2002, Vol 20, Issue 10, p1048
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt733