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Daniel Ehrlich

Publications and source records attributed to Daniel Ehrlich.

3 recordsLinked to original sources

Electrophoresis using ultra-high voltages.

Optimization of electrophoretic techniques is becoming an increasingly important area of research as microdevices are now routinely adapted for numerous biology and engineering applications. The present work seeks to optimize electrophoresis within microdevices by utilizing ultra-high voltages to increase sample concentration prior to separation. By imaging fluorescently-tagged DNA samples, the effects of both conventional and atypical voltage protocols on DNA migration and separation are readily observed. Experiments illustrate that short periods of high voltage during electrophoretic injection do not destroy the quality of DNA separations, and in fact can enhance sample concentration five-fold. This study presents data that illustrate increases in average resolution, and resolution of longer fragments, obtained from electrophoretic injections utilizing voltages between 85 and 850 V/cm.

DNA↗

Electrophoretic injection within microdevices.

The flexibility of the microfabricated format creates unique opportunities for study of the electrophoretic process. The present work utilizes digital images to capture the motion of DNA samples during pre-electrophoretic processes. A systematic study of DNA loading and strong sample stacking (sample concentration effects) was performed in order to analyze realistic DNA analysis conditions within microdevices. Using digital imaging and microscopy, DNA sample profiles within the injector were analyzed by deconvolving the geometrical intensity profile into different velocity groups. This analysis illustrates the evolution of molecular separation into distinct migrating populations within the injector itself. The present study performed DNA injections within microfabricated devices imposing run voltages between 85 and 850 V/cm. Data from 3 different offset lengths of a double-T cross-injector, 10 different applied voltages, and 2 different sample preparation protocols are presented.

DNA↗

High-speed analysis of multiplexed short tandem repeats with an electrophoretic microdevice.

We report the development of a robust and effective method for multiplexed short tandem repeat (STR) analysis within a chip-based microdevice. The method uses a laser-induced fluorescence detection system and simultaneously detects three- and four-color multiplexed polymerase chain reaction (PCR) samples. Analyses of the eight combined DNA index system (CODIS) STR loci were performed in 20 min with single-base-pair resolution ranging from 0.75 to 1. A simultaneous analysis of fifteen loci-ladders and a gender marker Amelogenin based on the PowerPlexTM 16 System was achieved in less than 35 min. The system is capable of repetitive operation and may be extended to high-throughput multilane devices that could be readily interfaced to an automated sample loading system.

Electrophoresis↗