Characterization of dye-induced mobility shifts affecting DNA sequencing in poly(ethylene oxide) sieving matrix.
Characterization of dye-induced mobility shifts affecting DNA sequencing in poly(ethylene oxide) sieving matrix.
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聚环氧乙烷筛分基质中染料引起的迁移率变化影响 DNA 测序的表征。
DOI：
10.1002/elps.1150181529
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Yeung,ES
中科院分区：
文献类型：
--
作者：
Tan,H;Yeung,ES
用激光诱导荧光检测和聚氧化乙烯填充毛细管电泳技术研究了三种荧光标记物（二吡咯甲撑二氟化硼（BODIPY）、能量转移（ET）和常规荧光素和罗丹明（ABI））对DNA测序的影响。在热空气热循环仪中通过染料标记的引物循环测序反应产生DNA测序片段。引入一个参数，相对诱导位移，以量化这些片段的电泳迁移率的均匀性。BODIPY被发现有最小的，但非零，染料诱导的不均匀性的影响。尽管ET染料由于其独特的光谱特性而提供了最高的灵敏度，但发现它们与BODIPY和ABI染料相比缺乏光稳定性。表征也带来了一些重要的提示，为选择合适的染料集的双通道比率为基础的DNA碱基调用方法。
The influence of three classes of fluorescence labels including dipyrrometheneboron difluoride (BODIPY), energy transfer (ET) and conventional fluorescein and rhodamine (ABI), on DNA sequencing has been examined with laserinduced fluorescence detection and poly (ethy1ene oxide)-filled capillary electrophoresis. DNA sequencing fragments were generated by dye-labeled primer cycle-sequencing reactions in a hot-air thermal cycler. A parameter, relativeinduced shift, was introduced to quantify the uniformity of electrophoretic mobilities of these fragments. BODIPY was found to have the smallest, but nonzero, effect for dye-induced nonuniformity. Although ET dyes provided the highest sensitivity due to their unique spectroscopic properties, they were found to lack photostability compared to BODIPY and ABI dyes. Characterization also brings out some important tips for selecting the suitable dye set for the two-channel ratio-based DNA base-calling method.