Chromatin Immunoprecipitation (IP) Fig 8-12

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5 Terms

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Step 1 - Experiment

  • live cultured cells or tissues incubated in 1% formaldehyde covalently cross-link proteins to DNA and proteins to proteins

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Step 2 - Experiment

  • preparation sonicated to solubilize chromatin and shear it into 200-500 base pair fragments of DNA

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Step 3 - Experiment

  • anti-RNA polymerase II antibody precipitates RNA polymerase II and its covalently linked DNA

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Step 4 - Experiment

  • reverse cross-linking, and isolate DNA

  • sequence DNA by massively parallel DNA sequencing

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What is the Purpose?

  • Chromatin immunoprecipitation using an antibody to RNA polymerase II: evidence for divergent transcription from most CpG island promoters in mammals.