Real-time fluorescence lifetime determinations for DNA sequencing
Abstract
Real-time determination of fluorescence lifetimes is investigated for improved identification of DNA bands during sequencing. On-the-fly identification is made possible through selectivity based in the time-domain. This method could involve using a set of fluorescent tags, 1-pyrene butanoic acid and increasingly substituted derivatives thereof, which possess varying lifetimes. These tags are covalently attached to primer strands and then selectively incorporated into complementary DNA fragments which are produced via established dideoxy sequencing protocol. Accomplishments have included the development of a laser-based instrument which is capable of real-time determination of fluorescence lifetimes for the identification of fluorescent analytes as they elute during HPLC. This instrumentation is also capable of collecting entire fluorescence decays on-the-fly and then mathematically determining the individual lifetimes for multi-component peaks.
Degree
Ph.D.
Advisors
Lytle, Purdue University.
Subject Area
Analytical chemistry|Biochemistry
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