5-Propargylamino-3'-azidomethyl-dCTP

产品基本信息

产品名称：5-Propargylamino-3'-azidomethyl-dCTP

储存条件：-15℃避光防潮

保质期：24个月

产品物理化学光谱特性

分子量：575.26

溶剂：水

产品介绍

5-Propargylamino-3'-azidomethyl-dCTP是制备用于下一代测序（NGS）的荧光缀合物的关键组成部分。 NGS使用与早期Sanger测序相似的链终止方法，但NGS是通过荧光标记的核苷酸类似物作为扩增反应的可逆终止剂来进行的。 NGS依赖于可逆的DNA聚合阻断，而Sanger测序则使用ddNTP不可逆转的DNA聚合阻断。 NGS的另一个不同特征是通过桥式PCR进行体外克隆扩增以增加待测序分子的数量。在此平台上，片段与固定在固体表面上的引物连接，进行原位扩增，生成具有相同分子的DNA簇。在每个循环中，同时添加可逆终止的四个核苷酸，并通过它们互补的聚合酶掺入。通过将3'-OH基团替换为3'-o-叠氮基甲基，这些核苷酸被化学封闭，以防止聚合酶在每个循环中掺入一个以上的核苷酸。掺入核苷酸后，在不同通道中针对不同碱基测量荧光信号。关于下一循环，洗涤未掺入的核苷酸，并用TCEP去除3'端的化学封锁。一旦收集到荧光信号，就会开始一个新的循环，重复此动态过程，直到完成每个片段的测序为止。总之，NGS测序反应分三个步骤进行：核苷酸的添加，成像和通过荧光团裂解的3'-OH再生。

参考文献

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