title: A device for extraction, manipulation and stretching of DNA from single human chromosomes.
abstract: We describe the structure and operation of a micro/nanofluidic device in which individual metaphase chromosomes can be isolated and processed without being displaced during exchange of reagents. The change in chromosome morphology as a result of introducing protease into the device was observed by time-lapse imaging; pressure-driven flow was then used to shunt the chromosomal DNA package into a nanoslit. A long linear DNA strand (>1.3 Mbp) was seen to stretch out from the DNA package and along the length of the nanoslit. Delivery of DNA in its native metaphase chromosome package as well as the microfluidic environment prevented DNA from shearing and will be important for preparing ultra-long lengths of DNA for nanofluidic analysis.
authors.affiliation: DTU Nanotech, Oersted Plads, Technical University of Denmark, Kongens Lyngby, Denmark.
authors.forename: Kristian H
authors.initials: KH
authors.lastname: Rasmussen
authors.forename: Rodolphe
authors.initials: R
authors.lastname: Marie
authors.forename: Jacob M
authors.initials: JM
authors.lastname: Lange
authors.forename: Winnie E
authors.initials: WE
authors.lastname: Svendsen
authors.forename: Anders
authors.initials: A
authors.lastname: Kristensen
authors.forename: Kalim U
authors.initials: KU
authors.lastname: Mir
mesh_terms: D002877:Chromosomes, Human; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004058:Diffusion; D004548:Elasticity; D004867:Equipment Design; D006801:Humans; D008677:Metaphase; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D036103:Nanotechnology / Q000295:instrumentation*; D010447:Peptide Hydrolases / Q000378:metabolism
chemical_list: D004247:DNA; D010447:Peptide Hydrolases