通过使用TOTO和YOYO嵌入染料与激光诱导荧光——毛细管电泳增强PCR产物的检测。

Enhanced detection of PCR products through use of TOTO and YOYO intercalating dyes with laser induced fluorescence--capillary electrophoresis.

作者信息

Srinivasan K, Morris S C, Girard J E, Kline M C, Reeder D J

机构信息

Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899.

出版信息

Appl Theor Electrophor. 1993;3(5):235-9.

PMID:8218477

Abstract

Recent developments in the chemical synthesis of DNA-binding dyes have enhanced detection of polymerase chain reaction (PCR) products by capillary electrophoresis. These dyes are dimers of thiazole orange (TOTO) or oxazole orange (YOYO) and have a very high binding affinity for DNA (Haugland, 1992). These dyes show enhanced fluorescence signals when they bind to double-stranded DNA and their fluorescence in the unbound state is almost zero, making them extremely useful in detecting minute (fg) quantities of DNA. We report here the utility of these dyes in DNA typing applications using a laser-induced fluorescence detector in conjunction with a capillary electrophoresis system.

摘要

DNA结合染料化学合成方面的最新进展增强了通过毛细管电泳对聚合酶链反应（PCR）产物的检测。这些染料是噻唑橙（TOTO）或恶唑橙（YOYO）的二聚体，对DNA具有非常高的结合亲和力（豪格兰德，1992年）。这些染料与双链DNA结合时会显示增强的荧光信号，且其未结合状态下的荧光几乎为零，这使得它们在检测微量（fg）DNA时极为有用。我们在此报告这些染料在使用激光诱导荧光检测器结合毛细管电泳系统的DNA分型应用中的效用。