Nonradioactive labeling of RNA
Nonradioactive labeling of RNA
复制标题
DOI：
10.1006/abio.1996.0016
复制
发表时间：
1996-01-01
影响因子：
2.9
通讯作者：
Igloi, GL
中科院分区：
文献类型：
--
作者：
Igloi, GL
利用DNA合成器上的商业成分合成了一种修饰的核二核苷酸(命名为pSEEp)。3‘端伯氨基的存在允许其与一系列非放射性标记物偶联，这里以荧光素为例。在非常温和的条件下，亲本二核苷酸的化学衍生化产物是T4RNA连接酶介导的RNA偶联的良好底物；靶RNA本身不经过化学处理。这种末端标记的明确产物可以通过例如使用基于荧光的DNA测序仪来分析。文件中的应用证明了这种方法的力量，这表明任何依赖于RNA检测或靶向标记的程序都可以适应于使用pSEEp及其衍生物。(C)1996年学术出版社。
A modified ribodinucleotide (named pSEEp) has been synthesized using commercially available components on a DNA synthesizer. The presence of a 3'-terminal primary amino group permits its coupling to a range of nonradioactive labels, exemplified here by fluorescein. The product of chemical derivatization of the parent dinucleotide is a good substrate for T4 RNA ligase-mediated coupling to RNA under very mild conditions; the target RNA is not itself subjected to chemical treatment. The well-defined product of this terminal labeling may be analyzed by, for example, the use of a fluorescence-based DNA sequencer. The applications documented serve to demonstrate the power of this approach suggesting that any procedure depending on the detection or targeted tagging of RNA may be adapted to using pSEEp and its derivatives. (C) 1996 Academic Press, Inc.