Sood Anup, Kumar Shiv, Nampalli Satyam, Nelson John R, Macklin John, Fuller Carl W
GE Healthcare, 800 Centennial Avenue, Piscataway, New Jersey 08855, USA.
J Am Chem Soc. 2005 Mar 2;127(8):2394-5. doi: 10.1021/ja043595x.
Nucleotides with a dye attached to the terminal phosphate with four or more phosphates (tetra- or pentaphosphates) are superior substrates than the corresponding triphosphates for DNA and RNA polymerases. When fluorogenic dyes are directly attached to the terminal phosphate, they can be released by the action of polymerase and alkaline phosphatase. The released dye changes color and fluorescence properties. The fluorescent signal can also be amplified by using multiple labeled nucleotides to detect small amounts of template. We have explored the utility of these nucleotides in a variety of applications including homogeneous SNP detection methods, DNA sequencing, and quantitation of PCR and RCA.
带有连接至末端磷酸基团（具有四个或更多磷酸基团，即四磷酸或五磷酸）的染料的核苷酸，对于DNA和RNA聚合酶而言，是比相应三磷酸核苷酸更好的底物。当荧光染料直接连接至末端磷酸基团时，它们可通过聚合酶和碱性磷酸酶的作用而被释放。释放出的染料会改变颜色和荧光特性。荧光信号也可通过使用多个标记的核苷酸来放大，以检测少量模板。我们已经探索了这些核苷酸在多种应用中的效用，包括均相单核苷酸多态性检测方法、DNA测序以及聚合酶链反应和滚环扩增的定量分析。
