DNA fragment sizing and sorting by laser-induced fluorescence
Abstract
A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.
- Inventors:
-
- Angier, NC
- Los Alamos, NM
- Issue Date:
- Research Org.:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- OSTI Identifier:
- 870612
- Patent Number(s):
- 5558998
- Assignee:
- Regents of Univ. of California (Alameda, CA)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- DOE Contract Number:
- W-7405-ENG-36
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- dna; fragment; sizing; sorting; laser-induced; fluorescence; method; provided; fragments; speed; detection; systems; flow; cytometry; determine; unique; characteristics; pieces; sample; characterization; piece; fragmented; preselected; sites; produce; plurality; resulting; treated; dye; effective; stain; stoichiometrically; stained; examined; generate; output; functionally; related; nucleotides; embodiment; intensity; emissions; linearly; length; distribution; sizes; forms; forensic; applications; laser-induced fluorescence; detection systems; functionally related; dna fragments; fluorescence emission; flow cytometry; dna fragment; unique characteristics; induced fluorescence; fluorescence emissions; linearly related; fragment sizing; unique characteristic; /435/436/
Citation Formats
Hammond, Mark L, Jett, James H, Keller, Richard A, Marrone, Babetta L, and Martin, John C. DNA fragment sizing and sorting by laser-induced fluorescence. United States: N. p., 1996.
Web.
Hammond, Mark L, Jett, James H, Keller, Richard A, Marrone, Babetta L, & Martin, John C. DNA fragment sizing and sorting by laser-induced fluorescence. United States.
Hammond, Mark L, Jett, James H, Keller, Richard A, Marrone, Babetta L, and Martin, John C. Mon .
"DNA fragment sizing and sorting by laser-induced fluorescence". United States. https://www.osti.gov/servlets/purl/870612.
@article{osti_870612,
title = {DNA fragment sizing and sorting by laser-induced fluorescence},
author = {Hammond, Mark L and Jett, James H and Keller, Richard A and Marrone, Babetta L and Martin, John C},
abstractNote = {A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1996},
month = {1}
}
Works referenced in this record:
Detection of single fluorescent molecules
journal, November 1990
- Brooks Shera, E.; Seitzinger, Newton K.; Davis, Lloyd M.
- Chemical Physics Letters, Vol. 174, Issue 6
Chromosome measurement and sorting by flow systems.
journal, April 1975
- Gray, J. W.; Carrano, A. V.; Steinmetz, L. L.
- Proceedings of the National Academy of Sciences, Vol. 72, Issue 4
A fluorescent complex between ethidium bromide and nucleic acids
journal, July 1967
- Lepecq, J. -B.; Paoletti, C.
- Journal of Molecular Biology, Vol. 27, Issue 1
Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications
journal, January 1992
- Rye, Hays S.; Yue, Stephen; Wemmer, David E.
- Nucleic Acids Research, Vol. 20, Issue 11
High-speed chromosome sorting
journal, October 1987
- Gray, J.; Dean, P.; Fuscoe, J.
- Science, Vol. 238, Issue 4825
A high-resolution, fluorescence-based, semiautomated method for DNA fingerprinting
journal, February 1989
- Carrano, A. V.; Lamerdin, J.; Ashworth, L. K.
- Genomics, Vol. 4, Issue 2
Single-molecule detection of Rhodamine 6G in ethanolic solutions using continuous wave laser excitation
journal, March 1991
- Soper, Steven A.; Shera, E. Brooks.; Martin, John C.
- Analytical Chemistry, Vol. 63, Issue 5
Rapid sizing of individual fluorescently stained DNA fragments by flow cytometry
journal, January 1993
- Goodwin, Peter M.; Johnson, Mitchell E.; Martin, John C.
- Nucleic Acids Research, Vol. 21, Issue 4
DNA Profiling: New tool links evidence to suspects with high certainty
journal, November 1989
- Thornton, John I.
- Chemical & Engineering News Archive, Vol. 67, Issue 47
Fluorescence polarization and pulse width analysis of chromosomes by a flow system.
journal, January 1979
- Cram, L. S.; Arndt-Jovin, D. J.; Grimwade, B. G.
- Journal of Histochemistry & Cytochemistry, Vol. 27, Issue 1
Chapter 37 Fluorescence in Situ Hybridization with DNA Probes
book, January 1990
- Trask, Barbara; Pinkel, Dan
- Flow Cytometry
High resolution chromosome analysis: one and two parameter flow cytometry
journal, January 1979
- Gray, J. W.; Langlois, R. G.; Carrano, A. V.
- Chromosoma, Vol. 73, Issue 1
Laser-Induced Fluorescence Detection of Rhodamine-6G at 6 × 10 −15 M
journal, June 1991
- Hahn, Jong Hoon; Soper, Steven A.; Nutter, Harvey L.
- Applied Spectroscopy, Vol. 45, Issue 5