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Proceedings Paper

Rapid sequencing of DNA based on single-molecule detection
Author(s): Steven A. Soper; Lloyd M. Davis; Frederick R. Fairfield; Mark L. Hammond; Carol A. Harger; James H. Jett; Richard A. Keller; Babetta L. Marrone; John C. Martin; Harvey L. Nutter; E. Brooks Shera; Daniel J. Simpson
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Paper Abstract

Sequencing the human genome is a major undertaking considering the large number of nucleotides present in the genome and the slow methods currently available to perform the task. The authors have recently reported on a scheme to sequence DNA rapidly using a non-gel based technique. The concept is based upon the incorporation of fluorescently labeled nucleotides into a strand of DNA, isolation and manipulation of a labeled DNA fragment and the detection of single nucleotides using ultra-sensitive laser-induced fluorescence detection following their cleavage from the fragment. Detection of individual fluorophores in the liquid phase was accomplished with time-gated detection following pulsed-laser excitation. The photon bursts from individual rhodamine 6G (R6G) molecules travelling through a laser beam have been observed, as have bursts from single fluorescently modified nucleotides. Using two different biotinylated nucleotides as a model system for fluorescently labeled nucleotides, the authors have observed synthesis of the complementary copy of M13 bacteriophage. Work with fluorescently labeled nucleotides is underway. Individual molecules of DNA attached to a microbead have been observed and manipulated with an epifluorescence microscope.

Paper Details

Date Published: 1 July 1991
PDF: 11 pages
Proc. SPIE 1435, Optical Methods for Ultrasensitive Detection and Analysis: Techniques and Applications, (1 July 1991); doi: 10.1117/12.44241
Show Author Affiliations
Steven A. Soper, Los Alamos National Lab. (United States)
Lloyd M. Davis, Los Alamos National Lab. (United States)
Frederick R. Fairfield, Los Alamos National Lab. (United States)
Mark L. Hammond, Los Alamos National Lab. (United States)
Carol A. Harger, Los Alamos National Lab. (United States)
James H. Jett, Los Alamos National Lab. (United States)
Richard A. Keller, Los Alamos National Lab. (United States)
Babetta L. Marrone, Los Alamos National Lab. (United States)
John C. Martin, Los Alamos National Lab. (United States)
Harvey L. Nutter, Los Alamos National Lab. (United States)
E. Brooks Shera, Los Alamos National Lab. (United States)
Daniel J. Simpson, Los Alamos National Lab. (United States)


Published in SPIE Proceedings Vol. 1435:
Optical Methods for Ultrasensitive Detection and Analysis: Techniques and Applications
Bryan L. Fearey, Editor(s)

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