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Isolation of Active Caenorhabditis elegans Nuclear Extract and Reconstitution for In Vitro Transcription
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生物化学
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JoVE 杂志 生物化学
Isolation of Active Caenorhabditis elegans Nuclear Extract and Reconstitution for In Vitro Transcription

Isolation of Active Caenorhabditis elegans Nuclear Extract and Reconstitution for In Vitro Transcription

DOI:
10.3791/62723-v

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06:09 min

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August 11, 2021

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1Department of Translational Medicine and Physiology, Elson S. Floyd College of Medicine,Washington State University

Chapters

  • 00:05Introduction
  • 00:23Animal and Balch Homogenizer Preparation
  • 01:41Animal Collection
  • 02:21Fractionation
  • 04:57Results: Quality Assessment of Nuclear Extract
  • 05:27Conclusion

Summary

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Here, we describe a detailed protocol for isolating active nuclear extract from larval stage 4 C. elegans and visualizing transcription activity in an in vitro system.

Tags

Caenorhabditis ElegansNuclear ExtractIn Vitro TranscriptionBalch HomogenizerHypotonic BufferHypertonic BufferSynchronizationL1 AnimalsL4 StageOP50CentrifugationHomogenizationTungsten Carbide Ball

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