STABLE FLUORESCENT COMPLEXES OF DOUBLE-STRANDED DNA WITH BIS-INTERCALATING ASYMMETRIC CYANINE DYES - PROPERTIES AND APPLICATIONS
STABLE FLUORESCENT COMPLEXES OF DOUBLE-STRANDED DNA WITH BIS-INTERCALATING ASYMMETRIC CYANINE DYES - PROPERTIES AND APPLICATIONS
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DOI：
10.1093/nar/20.11.2803
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发表时间：
1992-06-11
影响因子：
14.9
通讯作者：
GLAZER, AN
中科院分区：
文献类型：
--
作者：
RYE, HS;YUE, S;GLAZER, AN
报道了两种新的不对称菁染料噻唑橙二聚体（TOTO; 1,1′-（4,4,7,7-四甲基-4,7-重氮十一亚甲基）-双-4-[3-甲基-2,3-二氢-（苯并1,3-噻唑）-2-甲基二烯]-四碘喹啉二聚体）和恶唑黄二聚体（YOYO；用苯并1,3-恶唑代替苯并1,3-噻唑的TOTO类似物）的合成、结构证明、吸收和荧光性质。TOTO和YOYO在溶液中几乎无荧光，但与双链DNA （dsDNA）形成高荧光复合物，染料与DNA的最大bp比高达1:4，荧光增强达1000倍。ddna - toto （lambda(max) 513 nm）和ddna - yoyo (lambda(max) 489 nm)；λ (max)F 509 nm)配合物在琼脂糖和丙烯酰胺凝胶上电泳完全稳定。用乙锭二聚体（EthD; lambda(max)F 616 nm）预标记的限制性内切酶片段与TOTO或YOYO预标记的限制性内切酶片段的混合物通过电泳分离。488nm激光激发和同时共聚焦荧光检测在620 - 750 nm （dsDNA-EthD发射）和500 - 565 nm （dsDNA-TOTO或dsDNA-YOYO发射）允许灵敏的检测，定量和精确的大小限制片段从600到24000 bp。激光激发共聚焦荧光凝胶扫描仪在1毫米厚琼脂糖凝胶上5毫米宽的条带上检测dsDNA-TOTO和YOYO复合物的限限为4皮克，比传统的溴化乙啶染色低约500倍。
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.