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Chromatin Immunoprecipitation in the Cnidarian Model System Exaiptasia diaphana
JoVE Journal
Genetica
Author Produced
This content is Free Access.
JoVE Journal Genetica
Chromatin Immunoprecipitation in the Cnidarian Model System Exaiptasia diaphana
DOI:

11:48 min

March 17, 2023

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Capitoli

  • 00:00Introduction
  • 00:30Protein&#8209DNA Cross&#8209Linking
  • 02:13Homogenization and Lysis
  • 03:30Sonication
  • 04:08Immunoprecipitation (IP)
  • 05:28Recovery of the Immunocomplexes with Magnetic Beads
  • 06:20Washing, Elution, and Cross&#8209Linking Reversal
  • 08:13Protein and RNA Digestion and DNA Purification
  • 10:07Results: The Specificity and Effectiveness of the Assay by Quantitative PCR and Next&#8209Generation Sequencing
  • 11:28Conclusion

Summary

Traduzione automatica

A chromatin immunoprecipitation (X-ChIP) assay against the histone mark H3K4me3 for the model organism Exaiptasia diaphana is presented. The specificity and effectiveness of the assay are confirmed by quantitative PCR and next-generation sequencing. This protocol enables the increased investigation of protein-DNA interactions in the sea anemone E. diaphana.

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