Actions for Ultrasensitive fluorescence detection of DNA. Progress report, February 1, 1991--January 31, 1992 [electronic resource].

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Ultrasensitive fluorescence detection of DNA. Progress report, February 1, 1991--January 31, 1992 [electronic resource].

Published
Washington, D.C. : United States. Dept. of Energy, 1992.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy.
Physical Description
7 pages : digital, PDF file
Additional Creators
University of California, Berkeley, United States. Department of Energy, and United States. Department of Energy. Office of Scientific and Technical Information
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Summary
We have shown that a number of polycationic highly fluorescent dyes form complexes with double-stranded DNA (dsDNA) which are stable to electrophoresis and have characterized in detail such dsDNA complexes with TOTO (1,1′-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiazole)-2-methylidene]-quinolinium tetraiodide) and oxazole yellow dimer (YOYO; an analogue of TOTO with a benzo-1,3-oxazole in place of the benzo-1,3-thiazole). TOTO and YOYO are virtually non-fluorescent in solution, but form highly fluorescent complexes with dsDNA, up to a maximum dye to DNA bp ratio of 1:4, with >1000-fold fluorescence enhancement. We have developed an assay using YOYO for the quantitation of single-stranded and dsDNA in solution applicable over a range of DNA concentrations from 0.5 to 100 ng per ml. The fluorescent dsDNA-dye complexes allow detection of dsDNA on agarose and acrylamide gels with picogram sensitivity. We have applied these complexes in multiplex mapping experiments for accurate sizing and quantitation of restriction fragments. We have shown that in gel shift experiments the stable dsDNA-dye complexes can be used to detect heteroduplex-Muts complexes with a sensitivity comparable to radioisotopic detection.
Report Numbers
E 1.99:doe/er/61125--2
doe/er/61125--2
Subject(s)
Other Subject(s)
Note
Published through SciTech Connect.
12/01/1992.
"doe/er/61125--2"
"DE93004145"
Mathies, R.A.; Glazer, A.N.
Type of Report and Period Covered Note
Annual; 01/01/1991 - 12/31/1992
Funding Information
FG03-91ER61125
View MARC record | catkey: 13812035