title: A microfluidic system for large DNA molecule arrays.
abstract: Single molecule approaches offer the promise of large, exquisitely miniature ensembles for the generation of equally large data sets. Although microfluidic devices have previously been designed to manipulate single DNA molecules, many of the functionalities they embody are not applicable to very large DNA molecules, normally extracted from cells. Importantly, such microfluidic devices must work within an integrated system to enable high-throughput biological or biochemical analysis-a key measure of any device aimed at the chemical/biological interface and required if large data sets are to be created for subsequent analysis. The challenge here was to design an integrated microfluidic device to control the deposition or elongation of large DNA molecules (up to millimeters in length), which would serve as a general platform for biological/biochemical analysis to function within an integrated system that included massively parallel data collection and analysis. The approach we took was to use replica molding to construct silastic devices to consistently deposit oriented, elongated DNA molecules onto charged surfaces, creating massive single molecule arrays, which we analyzed for both physical and biochemical insights within an integrated environment that created large data sets. The overall efficacy of this approach was demonstrated by the restriction enzyme mapping and identification of single human genomic DNA molecules.
authors.affiliation: Laboratory for Molecular and Computational Genomics, Department of Chemistry, and Laboratory of Genetics, University of Wisconsin-Madison, 425 Henry Mall, Madison, Wisconsin 53706, USA.
authors.forename: Eileen T
authors.initials: ET
authors.lastname: Dimalanta
authors.forename: Alex
authors.initials: A
authors.lastname: Lim
authors.forename: Rod
authors.initials: R
authors.lastname: Runnheim
authors.forename: Casey
authors.initials: C
authors.lastname: Lamers
authors.forename: Chris
authors.initials: C
authors.lastname: Churas
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Forrest
authors.forename: Juan J
authors.initials: JJ
authors.lastname: de Pablo
authors.forename: Michael D
authors.initials: MD
authors.lastname: Graham
authors.forename: Susan N
authors.initials: SN
authors.lastname: Coppersmith
authors.forename: Steve
authors.initials: S
authors.lastname: Goldstein
authors.forename: David C
authors.initials: DC
authors.lastname: Schwartz
mesh_terms: D004247:DNA / Q000737:chemistry*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008970:Molecular Weight; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods
chemical_list: D004247:DNA