Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

YingWu; Ya Ying Zheng; Qishan Lin

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Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

机译：检测和量化的RNAPhosphorothioate修改使用质量光谱法

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This article describes a protocol for detecting and quantifying RNA phosphorothioate modifications in cellular RNA samples. Starting from solid-phase synthesis of phosphorothioate RNA dinucleotides, followed by purification with reversed-phase HPLC, phosphorothioate RNA dinucleotide standards are prepared for UPLC-MS and LC-MS/MS methods. RNA samples are extracted from cells using TRIzol reagent, then digested with a nuclease mixture and analyzed by mass spectrometry. UPLC-MS is employed first to identify RNA phosphorothioate modifications. An optimized LC-MS/MS method is then employed to quantify the frequency of RNA phosphorothioate modifications in a series of model cells.

机译：本文介绍了一种检测的协议和量化RNA phosphorothioate修改在细胞RNA样本。phosphorothioate固相合成RNA二核苷酸,其次是净化反相高效液相色谱法,phosphorothioate RNA二核苷酸UPLC-MS标准准备和质/ MS方法。使用试剂盒从细胞试剂,然后消化核酸酶混合物和分析质量谱分析。识别RNA phosphorothioate修改。然后采用优化的质/ MS方法量化RNA phosphorothioate的频率修改在一系列的细胞模型。

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《Current Protocols in Nucleic Acid Chemistry》 |2020年第1期|e113/1-e113/11|共11页
作者
YingWu; Ya Ying Zheng; Qishan Lin;
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作者单位

Department of Chemistry and The RNA Institute, University at AlbanyState University of New York, Albany, New York;

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原文格式 PDF
正文语种英语
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关键词
MODIFICATION; Nucleic acids; thiophosphoric acidSolid-Phase SynthesisRNA;

机译：修改;核酸;thiophosphoricacidSolid-Phase SynthesisRNA;

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Detection and Quantification of RNA Phosphorothioate Modifications Using Mass Spectrometry

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