The goal of this project is to develop a new data acquisition and information processing system for electrophoretic DNA sequencing applications, such that there will be no need for human intervention all the way to the final base-pair calling. The system output will be accompanied by a quantitative confidence measure for each sequence call. The proposed system is essentially based on a new technique, based on a multicolor illumination of the analyte by several semiconductor lasers, tuned to th absorption peaks of different fluorophore labels. Each laser will be modulated in intensity at a distinct radio-frequency and the received fluorescent signal will be frequency analyzed in the electric domain. This technique provides additional information about the signal, permitting a significant enhancement of the confidence level in base pair identification and enabling a fully automated procedure without human experts. Practicality of this technique is based on the use of cheap and miniature semiconductor lasers, which permit an easy modulation of output radiation at a well defined radio frequency. The use of amplitude modulation and frequency detection has been shown to enhance the signal to noise ratio by at least a factor of 10. The miniature size enables the installation of individual read-out lines on each electrophoretic lane or capillary. This approach admits of practically unlimited parallelism. Estimates shown that the cost of data acquisition will be essentially eliminated from the budget consideration. The system will be primarily oriented on the use of infrared dyes, although its modular architecture and novel fiber-optical probe could be adapted to any wavelength used for exciting fluorescence. Five pilot prototype systems will be produced in the second ad third year of the project. The prototypes will be adapted to slab-gel capillary, ultra-thin gel and membrane systems. The characterization will be carried out both at Stony Brook and in collaboration with our industrial partners.
| Bilenko, Olga; Gavrilov, Dmitri; Gorbovitski, Boris et al. (2003) Formation of a resistive region at the anode end in DNA capillary electrophoresis. Electrophoresis 24:1176-83 |
