Quantitative analysis of RNA modifications.
Quantitative analysis of RNA modifications.
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DOI：
10.1007/978-1-60761-657-3_2
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Yu, Yi-Tao
中科院分区：
文献类型：
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作者：
Karijolich, John;Kantartzis, Athena;Yu, Yi-Tao
RNA 修饰影响许多细胞过程，包括前 mRNA 剪接和蛋白质合成。阐明这些修饰影响细胞过程的机制需要能够检测和量化 RNA 中这些修饰的存在。在这里，我们提出了一个详细的程序，可以检测和定量 RNA 碱基修饰。该过程涉及多种技术，包括寡核苷酸亲和选择、位点特异性切割和放射性标记、核酸酶消化和薄层色谱。
RNA modifications impact numerous cellular processes including pre-mRNA splicing and protein synthesis. The elucidation of the mechanisms by which these modifications impact cellular processes necessitates the ability to both detect and quantify the presence of these modifications within RNA. Here, we present a detailed procedure that allows for both detecting and quantifying RNA base modifications. This procedure involves a number of techniques, including oligonucleotide-affinity selection, site-specific cleavage and radiolabeling, nuclease digestion, and thin layer chromatography.