The goal of this project is to develop microfabricated devices for the preparation and electrophoretic separation of DNA for the purposes of high-throughput, rapid sequencing at low cost. This project will focus on three areas: I) compact micromachined channel structures for capillary gel electrophoresis with performance equivalent of exceeding that demonstrated for conventional capillaries, 2) integration of micro-reactors and mixers for production and clean-up of Sanger sequencing products on the electrophoresis chip, and 3) integration of a hydrodynamic focusing flow cell on the electrophoresis chip for ultrasensitive fluorescence detection of dye end-labeled Sanger products. The advantages of microfabrication and integration of functionality are many: I) the ability to manipulate small volumes (10pL - 10nL) of fluids with high precision (< 1% rsd) on microchips, 2) reduction in reagent volume by as much as 10(4), 3) automation of chemical procedures with no moving parts at low cost, 4) massively parallel operations with minimal space and cost requirements. The dexterity of the fluidic manipulations that we have previously developed encourages loading of the sample onto the chip as early in the process as possible. Success of this project will foster further integration of chemical processing on microchips back to the stage of clone picking. Such microdevices could be directly inserted into shotgun or directed sequencing programs.
| Khandurina, J; Jacobson, S C; Waters, L C et al. (1999) Microfabricated porous membrane structure for sample concentration and electrophoretic analysis. Anal Chem 71:1815-9 |
