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高分辨率基因表达谱RNA的合成，加工和衰变的新转录RNA在细胞培养的代谢标记
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Biologia
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JoVE Journal Biologia
Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
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高分辨率基因表达谱RNA的合成，加工和衰变的新转录RNA在细胞培养的代谢标记

DOI:
10.3791/50195-v

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11:00 min

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August 08, 2013

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, , , , ,
1Max von Pettenkofer Institute, 2Department of Medicine,University of Cambridge, 3Institute for Informatics,Ludwig-Maximilians-University Munich

Capítulos

  • 00:05Título
  • 01:44Metabolic Labeling with 4-thiouridine
  • 03:05Thiol-specific Biotinylation of Newly Transcribed RNA
  • 05:09Separation of Labeled and Unlabeled RNA Using Streptavidin-coated Magnetic Beads
  • 07:00Recovery of Newly Transcribed RNA
  • 08:17Results: Metabolic Labeling of Newly Transcribed RNA
  • 10:25Conclusion

Summary

Tadução automática

Tadução automática

细胞总RNA提供了一种用于研究的短期变化，以及在RNA合成和衰变动力学的RNA加工模板差。这里，我们描述遵循与4 - thiouridine的新转录的RNA的代谢标记通过硫醇特异性的生物素化和纯化的新转录的RNA，来克服这些限制。

Tags

Metabolic Labeling4-thiouridine4sU-taggingRNA SynthesisRNA ProcessingRNA DecayGene Expression ProfilingTranscriptomeKineticsComputational Modeling

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