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利用体外和细胞内形状研究小分子诱导的预 mrna 结构变化

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Genetica

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JoVE Journal Genetica

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

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利用体外和细胞内形状研究小分子诱导的预 mrna 结构变化

DOI:

10.3791/59021-v

•

11:58 min

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January 30, 2019

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Jingxin Wang, John Hammond, Kristen A. Johnson

¹Calibr,The Scripps Research Institute, ²Department of Integrative Structural and Computational Biology,The Scripps Research Institute

Capitoli

00:04Titolo
00:31³²P-labeled Primer Labeling for In Vitro Selective 2‘-hydroxyl Acylation Analyzed by Primer Extension (SHAPE)
03:44Small Molecule Binding and RNA 2‘-OH Modification
06:29Marker Preparation and Primer Extension
08:59PAGE Analysis
10:15Results of In Vitro and In-cell SHAPE for Analyzing Small Molecule Induced Pre-mRNA Structural Changes
11:02Conclusion

Summary

Traduzione automatica

通过引物扩展 (shape) 实验分析的体外和细胞内选择性 2 '-羟基酰化的详细方案, 以确定在存在 rna 靶向小分子的情况下, mrna 前序列的二级结构。本文中介绍。

Tags

Keywords: In Vitro SHAPE In-cell SHAPE RNA Secondary Structure Small Molecule Pre-mRNA Reverse Transcription Radioactive Labeling Acrylamide Gel Passive Elution

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