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

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

DOI:
10.3791/59021-v

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11:58 min

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January 30, 2019

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

'Bölümler'

  • 00:04'Başlık'
  • 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

Özet

'Otomatik Çeviri'

'Otomatik Çeviri'

Detailed protocols for both in vitro and in-cell selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) experiments to determine the secondary structure of pre-mRNA sequences of interest in the presence of an RNA-targeting small molecule are presented in this article.

'Etiketler'

In Vitro SHAPEIn-cell SHAPERNA Secondary StructureSmall MoleculePre-mRNAReverse TranscriptionRadioactive LabelingAcrylamide GelPassive Elution

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