Quick two-step RNA ligation employing periodate oxidation
Quick two-step RNA ligation employing periodate oxidation
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DOI：
10.1093/nar/gng145
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发表时间：
2003-11-15
影响因子：
14.9
通讯作者：
Watanabe, K
中科院分区：
文献类型：
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作者：
Kurata, S;Ohtsuki, T;Watanabe, K
将修饰或标记的核苷酸引入RNA是一种强大的RNA工程工具，因为它使我们能够研究天然RNA修饰如何影响RNA的功能和结构。它也有助于RNA的结构分析。修饰的核苷酸可以通过两步酶促连接RNA片段的方法引入RNA的特定位置。然而，该方法在两个连接步骤之间需要复杂的纯化步骤，这导致连接产物的低产率。在这里，我们开发了一种新的连接技术，采用高碘酸氧化物，消除了这个纯化步骤。这增加了连接产物的总产率并使其成为更快的程序。
The introduction of modified or labeled nucleotides into RNA is a powerful RNA engineering tool as it enables us to investigate how native RNA modifications affect RNA function and structure. It also helps in the structural analysis of RNA. A modified nucleotide can be introduced into a specific position of RNA by the method of two-step enzymatic ligation of RNA fragments. However, this method requires a complicated purification step between the two ligation steps that results in low yields of the ligation product. Here we have developed a new ligation technique employing periodate oxide that eliminates this purification step. This increases the total yield of the ligation product and makes it a faster procedure.