Our goal is the development of an inexpensive automated 4-color single- lane DNA sequencing machine adapted for mutation diagnostics in clinical environment using the p53 gene as a model system. Operation of the machine is based on a recently disclosed multicolor fluorescent detection technique which enables the detection and analysis low-power fluorescence signals. Main features of the machine: . analysis of contaminated DNA samples (10% resolution of wild and mutated admixtures); . high throughput (up to 500,000 bp/year); . low amount of labeled DNA material (10 times stretched ABI sequencing kit); . low cost of the machine In Phase I in collaboration with the Department of Electrical Engineering, SUNY Stony Brook, we shall develop an ultra-sensitive 4-color fluorescent detection system with data transfer rate of 1.6 Mbit/s and, in collaboration with the Department of Pathology SUNY SB, test this detection system using clinical DNA material. In Phase II we shall develop a pilot prototype of the machine entirely based on semiconductor components, integrated technologies, and low-power illumination sources and validate the machine using the p53 gene as a model system. In Phase III we shall organize manufacturing of the single-capillary machine and develop a 16-lane sequencer for diagnostic applications.
The proposed automated sequencing machine will find applications in clinical diagnostics of genetic disease, primarily cancer. its unique capability to precisely quantify contaminated DNA samples will ensure a dominant position in an important market segment. The low cost, miniature 4-colored automated machine with low consumption of labeled DNA material will be attractive for individual researchers and medical diagnostic laboratories.
