Version 2.1.1.0

Abstract

Grant Number:	5R44HG002292-04
PI Name:	WILLIAMS, JOHN
PI Email:	jwilliams@licor.com
PI Title:	SENIOR SCIENTIST
Project Title:	DNA polymerase for single molecule DNA sequencing

Abstract: DESCRIPTION (provided by applicant): We are developing a new technology for single molecule DNA sequencing based on the incorporation of dNTPs labeled on the g-phosphate with fluorescent dyes. The label is cleaved from the nucleotide along with pyrophosphate when the nucleotide is incorporated into DNA by a polymerase. The released label is sorted from intact g-dNTPs in a microfluidics system and is identified in real time by single molecule fluorescence detection optics. In this proposal, we develop a novel nucleotide chemistry to enable the sorting scheme and we use directed evolution methods to adapt polymerases for utilization of the g-dNTPs. Single molecule sequencing promises to reduce the cost of DNA sequencing by several orders of magnitude compared to current methods. PROPOSED COMMERCIAL APPLICATIONS: A genomic DNA sequencing technology that reduces cost and eliminates the need for cloning, physical mapping and electrophoresis would have great appeal to both the academic and industrial genomic research communities.

Thesaurus Terms:
DNA directed DNA polymerase, biochemical evolution, high throughput technology, nucleic acid sequence, protein engineering, recombinant protein, technology /technique development
DNA footprinting, bacteriophage T7, fluorescent dye /probe, gene mutation, microarray technology, nucleotide analog
biotechnology, combinatorial chemistry, molecular cloning, nanotechnology, polymerase chain reaction, protein purification, site directed mutagenesis

Institution:	LI-COR, INC.
	4308 PROGRESSIVE AVE
	LINCOLN, NE 68504
Fiscal Year:	2003
Department:
Project Start:	30-SEP-2001
Project End:	31-JAN-2004
ICD:	NATIONAL HUMAN GENOME RESEARCH INSTITUTE
IRG:	ZRG1

---