Electromigration of single molecules of DNA in a crystalline array of 300-nm silica colloids.
The velocities and conformations of single DNA chains were probed as they electromigrated at varying electric field strength through a crystalline array of silica colloids. An optically transparent film consisting of 300-nm silica colloids was formed on glass as a transparent crystalline layer of 7 mum thickness with an effective pore size of 45 nm. The behaviors of individual lambda-DNA molecules (48,502 base pairs) electromigrating through this material were observed to be analogous to the behaviors of long DNA chains in electrophoresis gels, including chain extension, hooking of chains around the matrix, and hernia formation. The electrophoretic mobility of lambda-DNA in this dense, narrow-pore material is surprisingly high: 1.8 cm2/Vs at 10 V/cm, which is at least as high as for much wider-pore gels. Imaging of the single molecules revealed that higher field strength caused increased chain extension and increased mobility, which reached an apparent plateau just above 2.0 cm2/Vs at 200 V/cm. Pulsed, crossed electric fields of 200 V/cm at 120 degrees to one another were applied to the material. The DNA chains were observed by imaging to electromigrate in an orderly fashion, and the migration rate was found to be length-dependent. The results indicate that these thin, robust, self-assembling inorganic materials are interesting as possible alternatives to polymeric gels for higher speed electrophoresis.
Zhang
Hui
H
Department of Chemistry, 1306 East University Boulevard, University of Arizona, Tucson, Arizona 85721, USA.
Wirth
Mary J
MJ
D010582:Bacteriophage lambda / Q000737:chemistry; D003102:Colloids / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D004279:DNA, Viral / Q000737:chemistry; D053841:Electroosmosis; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods; D008855:Microscopy, Electron, Scanning; D012821:Silanes / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry*; D055672:Static Electricity
D003102:Colloids; D004279:DNA, Viral; D012821:Silanes; C040733:tetraethoxysilane; D012822:Silicon Dioxide; D004247:DNA