Abstract
Chromatin immunoprecipitation (ChIP) is an invaluable method to characterize interactions between proteins and genomic DNA, such as the genomic localization of transcription factors and posttranslational modification of histones. DNA and proteins are reversibly and covalently crosslinked using formaldehyde. Then the cells are lysed to release the chromatin. The chromatin is fragmented into smaller sizes either by micrococcal nuclease (MNase) or sonication and then purified from other cellular components. The protein-DNA complexes are enriched by immunoprecipitation (IP) with antibodies that target the epitope of interest. The DNA is released from the proteins by heat and protease treatment, followed by degradation of contaminating RNAs with RNase. The resulting DNA is analyzed using various methods, including PCR, qPCR, or sequencing. This protocol outlines each of these steps for both yeast and human cells.
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Acknowledgment
These protocols were developed with help from Aneeshkumar Arimbasseri and Kwan T. Chow. This work was supported by funds from North Carolina State University, NIH Grants R21E023377Â and 1DP1DA044359, the GI Bill, and Simons Foundation Grant 495112.
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Lee, J.B., Keung, A.J. (2018). Chromatin Immunoprecipitation in Human and Yeast Cells. In: Jeltsch, A., Rots, M. (eds) Epigenome Editing. Methods in Molecular Biology, vol 1767. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7774-1_14
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DOI: https://doi.org/10.1007/978-1-4939-7774-1_14
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