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用于分析依赖帽的翻译动力学的 体外 单分子成像测定
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Bioquímica
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JoVE Journal Bioquímica
An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
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用于分析依赖帽的翻译动力学的 体外 单分子成像测定

DOI:
10.3791/61648-v

•

09:52 min

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September 15, 2020

•

, ,
1Molecular Biology Program,Memorial Sloan Kettering Cancer Center

Capítulos

  • 00:05Introduction
  • 00:51Equipment Preparation
  • 03:113’-End Biotinylated mRNA Immobilization
  • 05:20Translation Mix Assembly and Delivery
  • 07:24Results: Analyses for Saccharomyces cerevisiae Translation of a Firefly Luciferase-encoding mRNA
  • 09:12Conclusion

Summary

Tadução automática

Tadução automática

跟踪单个翻译事件允许对依赖上限的翻译机制进行高分辨率动能研究。在这里，我们演示了基于荧光标记抗体和表皮标记的新生肽之间的成像相互作用的 体外 单分子检测。该方法使启动和肽伸长动力学的单分子特征 在主动体外 帽依赖翻译。

Tags

Keywords: Single-molecule ImagingCap-dependent TranslationIn Vitro TranslationCell-free TranslationMicroscope SetupLaser ControlCamera SettingsSyringe PumpWash Protocol

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