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1Red Sea Research Center (RSRC), Biological and Environmental Sciences and Engineering Division (BESE),King Abdullah University of Science and Technology (KAUST), 2KAUST Environmental Epigenetics Program (KEEP), Biological and Environmental Sciences and Engineering Division (BESE),King Abdullah University of Science and Technology (KAUST)
Chapters
- 00:00Introduction
- 00:30Protein‑DNA Cross‑Linking
- 02:13Homogenization and Lysis
- 03:30Sonication
- 04:08Immunoprecipitation (IP)
- 05:28Recovery of the Immunocomplexes with Magnetic Beads
- 06:20Washing, Elution, and Cross‑Linking Reversal
- 08:13Protein and RNA Digestion and DNA Purification
- 10:07Results: The Specificity and Effectiveness of the Assay by Quantitative PCR and Next‑Generation Sequencing
- 11:28Conclusion
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.
