微通道板毛细管电泳DNA分级的荧光标记方法。

Igor Medintz, Wendy W Wong, Richard A Mathies
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引用次数: 0

摘要

微通道板毛细管电泳平台上的DNA分析将随着荧光标记分析物和标准分级梯的发展而得到进一步的发展。在这里,我们评估了商用DNA阶梯的末端标记和末端脱氧核苷酸转移酶(TdT)的聚合酶链反应(PCR)扩增子作为荧光双链DNA大小分析的方法。一个基于聚合酶链反应的程序,方便建设自定义的能量转移标记的DNA分级阶梯也提出。单链DNA片段的高分辨率大小被证明与这个能量转移标记的梯子。这些DNA标记程序将对微设备和其他电泳平台上的双链和单链DNA分析有用。
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Fluorescence labeling methods for microchannel plate capillary electrophoresis DNA sizing.

DNA analysis on microchannel plate (MCP) capillary electrophoresis platforms will be advanced by the development of new methods for the fluorescence labeling of analytes and standard sizing ladders. Here we evaluate end-labeling of commercially available DNA ladders and polymerase chain reaction (PCR) amplicons with terminal deoxynucleotidyl transferase (TdT) as a method for fluorescent double-stranded DNA sizing analysis. A PCR-based procedure for the facile construction of custom energy-transfer-labeled DNA sizing ladders is also presented. High-resolution sizing of single-stranded DNA fragments is demonstrated with this energy-transfer-labeled ladder. These DNA labeling procedures will be useful for double-and single-stranded DNA analyses on microdevices and other electrophoretic platforms.

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