Splint ligation of RNA with T4 DNA ligase.
Splint ligation of RNA with T4 DNA ligase.
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DOI：
10.1007/978-1-62703-113-4_19
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
O'Keefe, Raymond T
中科院分区：
文献类型：
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作者：
Kershaw, Christopher J;O'Keefe, Raymond T
RNA的夹板连接，其中特定的RNA分子连接在一起，可以使用T4 DNA连接酶和与RNA互补的桥接DNA寡核苷酸进行。该方法利用T4 DNA连接酶的性质，当RNA分子处于RNA：DNA杂合体中时，将它们连接起来。夹板连接是将修饰的核苷酸引入RNA分子、将放射性标记插入RNA内的特定位置以及将较小的合成RNA组装成较长的RNA分子的有用工具。这样的修饰使得能够用修饰的RNA进行广泛的实验，包括结构研究、免疫共沉淀以及绘制RNA：RNA和RNA：蛋白质相互作用的位点的能力。
Splint ligation of RNA, whereby specific RNA molecules are ligated together, can be carried out using T4 DNA ligase and a bridging DNA oligonucleotide complementary to the RNAs. This method takes advantage of the property of T4 DNA ligase to join RNA molecules when they are in an RNA:DNA hybrid. Splint ligation is a useful tool for the introduction of modified nucleotides into RNA molecules, insertion of a radiolabel into a specific position within an RNA and for the assembly of smaller synthetic RNAs into longer RNA molecules. Such modifications enable a wide range of experiments to be carried out with the modified RNA including structural studies, co-immunoprecipitations, and the ability to map sites of RNA:RNA and RNA:protein interactions.