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| STABLE FLUORESCENT COMPLEXES OF DOUBLE-STRANDED DNA WITH BIS-INTERCALATING ASYMMETRIC CYANINE DYES - PROPERTIES AND APPLICATIONS |
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| Author(s): RYE HS, YUE S, WEMMER DE, QUESADA MA, HAUGLAND RP, MATHIES RA, GLAZER AN |
| Source: NUCLEIC ACIDS RESEARCH Volume: 20 Issue: 11 Pages: 2803-2812 Published: JUN 11 1992 |
| Times Cited: 343 References: 22 |
| Abstract: The synthesis, proof of structure, and the absorption and fluorescence properties of two new unsymmetrical cyanine dyes, thiazole orange dimer (TOTO; 1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiazole)-2-methyl-idene]-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) are reported. TOTO and YOYO are virtually non-fluorescent in solution, but form highly fluorescent complexes with double-stranded DNA (dsDNA), up to a maximum dye to DNA bp ratio of 1:4, with > 1 000-fold fluorescence enhancement. The dsDNA-TOTO (lambda(max) 513 nm lambda(max)F 532 nm) and dsDNA-YOYO (lambda(max) 489 n; lambda(max)F 509 nm) complexes are completely stable to electrophoresis on agarose and acrylamide gels. Mixtures of restriction fragments pre-labeled with ethidium dimer (EthD; lambda(max)F 616 nm) and those pre-labeled with either TOTO or YOYO were separated by electrophoresis. Laser excitation at 488 nm and simultaneous confocal fluorescence detection at 620 - 750 nm (dsDNA-EthD emission) and 500 - 565 nm (dsDNA-TOTO or dsDNA-YOYO emission) allowed sensitive detection, quantitation, and accurate sizing of restriction fragments ranging from 600 to 24,000 bp. The limit of detection of dsDNA-TOTO and YOYO complexes with a laser-excited confocal fluorescence gel scanner for a band 5-mm wide on a 1-mm thick agarose gel was 4 picograms, about 500-fold lower than attainable by conventional staining with ethidium bromide. |
| Document Type: Article |
| Language: English |
Addresses:
1. UNIV CALIF BERKELEY, DEPT MOLEC & CELL BIOL, MCB STANLEY DONNER ASU, 229 STANLEY HALL, BERKELEY, CA 94720 USA
2. UNIV CALIF BERKELEY, DEPT CHEM, BERKELEY, CA 94720 USA
3. LAWRENCE BERKELEY LAB, DIV CHEM BIODYNAM, BERKELEY, CA 94720 USA
4. MOLEC PROBES INC, EUGENE, OR 97402 USA |
| Publisher: OXFORD UNIV PRESS UNITED KINGDOM, WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP |
| Subject Category: Biochemistry & Molecular Biology |
| IDS Number: HZ673 |
| ISSN: 0305-1048 |
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