Effects of gel material on fluorescence lifetime detection of dyes and dye-labeled DNA primers in capillary electrophoresis.
Effects of gel material on fluorescence lifetime detection of dyes and dye-labeled DNA primers in capillary electrophoresis.
复制标题
凝胶材料对毛细管电泳中染料和染料标记 DNA 引物荧光寿命检测的影响。
DOI：
10.1016/s0021-9673(99)00292-7
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
McGown,LB
中科院分区：
文献类型：
--
作者：
Li,L;McGown,LB
染料6-[N-（2-甲基苯基）-1-甲基]-2-氧代苯胺荧光寿命的研究（7-硝基苯并-2-氧杂-1，3-二氮唑-4-基）氨基]己酸（NBD-HA）和用NBD-HA、Cy 3、罗丹明绿色和4，4-二氟-5，7-二甲基-4-硼杂-3a，4a-二氮杂-s-引达省-3-丙酸标记的DNA M13引物描述了在具有不同交联度和不同交联剂的聚丙烯酰胺凝胶中以及在具有不同有机改性剂的纤维素筛分缓冲液中的（BODIPY-FL）染料。凝胶基质和实验条件上的荧光寿命的依赖性进行了研究，以确定哪些因素可能是重要的优化多重荧光寿命检测。在批溶液和运行中、CE中的柱上测定寿命。结果表明，在不同的凝胶组成的引物连接的染料的寿命保持不变。此外，在纤维素筛分缓冲液的批处理溶液中观察到引物连接染料的多指数荧光衰减，但当在CE中进行实时柱上测量时，其减少为单指数衰减。通过向凝胶基质中加入有机改性剂可以提高寿命检测能力。
Investigations of fluorescence lifetimes of the dye 6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoic acid (NBD-HA) and of DNA M13 primers labeled with NBD-HA, Cy3, rhodamine green and 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid (BODIPY-FL) dyes in polyacrylamide gels of various degrees of crosslinking and different crosslinkers, and in a cellulose sieving buffer with different organic modifiers, are described. The dependence of fluorescence lifetime on gel matrix and on experimental conditions was studied in order to identify which factors may be important for optimization of multiplex fluorescence lifetime detection. Lifetimes were determined in both batch solution and on-the-fly, on-column in CE. Results show that lifetimes of the primer-attached dyes remain constant in gels of different composition. Additionally, multiexponential fluorescence decays are observed for primer-attached dyes in batch solutions of the cellulose sieving buffers but are reduced to monoexponential decays when measured on-the-fly, on-column in CE. Lifetime detectability can be improved by addition of an organic modifier to the gel matrix.