A DNA prism for high-speed continuous fractionation of large DNA molecules.
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.
Huang
Lotien Richard
LR
Center for Photonics and Optoelectronic Materials (POEM), Princeton University, Princeton, NJ 08544, USA.
Tegenfeldt
Jonas O
JO
Kraeft
Jessica J
JJ
Sturm
James C
JC
Austin
Robert H
RH
Cox
Edward C
EC
D005591:Chemical Fractionation / Q000295:instrumentation* / Q000379:methods*; D022202:Chromosomes, Artificial, Bacterial; D027042:Chromosomes, Artificial, P1 Bacteriophage; D004269:DNA, Bacterial / Q000032:analysis*; D004574:Electromagnetic Fields; D004586:Electrophoresis / Q000379:methods*; D036103:Nanotechnology; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*
D004269:DNA, Bacterial