Fluorescence Imaging Chip System for Massive Parallel DNA Sequencing

Columbia University, New York NY

Investigators

Abstract

0097793 Ju The objective of this research is the development of a fluorescence imaging system for massive parallel DNA sequencing. This sequencing system includes the construction of a chip with immobilized single stranded DNA templates that can self prime for the generation of the complementary DNA strand in the polymerase reaction. The system also includes 4 unique fluorescently labeled nucleotide analogues with capped by a small chemical moiety to allow efficient incorporation into the growing strand of DNA as terminators in the polymerase reaction. A 4-color fluorescence imager is then used to identify the sequence of the incorporated nucleotide on each spot of the chip. After removal of the dye photochemically and the capping group, the polymerase reaction proceeds to incorporate the next nucleotide analogue and detect the next base. High density (>10,000 spots per chip) single stranded DNA will be immobilized on a 4 cm. x 1 cm. glass chip in order to identify more than 10,000 bases after each cycle. After 100 cycles a million base pairs will be generated on one sequencing chip.