DNA electrophoresis in microlithographic arrays.

Autor: Volkmuth WD; Department of Physics, Princeton University, New Jersey 08544., Austin RH
Jazyk: angličtina
Zdroj: Nature [Nature] 1992 Aug 13; Vol. 358 (6387), pp. 600-2.
DOI: 10.1038/358600a0
Abstrakt: We have used optical microlithography to fabricate capped quasi-two-dimensional obstacle courses in SiO2. We report here observations using epifluorescence microscopy of the electrophoresis and length fractionation of large DNA molecules confined in arrays. Simple reptation theory, based on the work of deGennes, predicts that at low electric fields the electrophoretic mobility of a polymer of length L much greater than the persistence length p scales inversely with L (ref. 2). But elongation of the coil in the matrix at sufficiently strong electric fields results in a length-independent electrophoretic mobility. The application of suitably timed pulsed electric fields restores the fractionating power of gels for long molecules but the protocols of pulsed-field electrophoresis are semi-empirical because the complex and ill-understood gel matrix plays a critical role in fractionation. Microlithographically constructed obstacle arrays, with their low dimensionality, small volume and extremely reproducible topography, will make it possible to understand the motion and fractionation of large polymer molecules in complex but well characterized topologies.
Databáze: MEDLINE