Chapter title |
Topoisomerase II Chromatin Immunoprecipitation
|
---|---|
Chapter number | 14 |
Book title |
DNA Topoisomerases
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7459-7_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7458-0, 978-1-4939-7459-7
|
Authors |
Kayleigh A. Smith, Ian G. Cowell, Caroline A. Austin |
Abstract |
Chromatin immunoprecipitation is a method to isolate a protein of interest coupled to DNA following cross-linking with formaldehyde and to quantify the relative abundance or occupancy of the protein at specific genomic loci. After immunoprecipitation of protein-DNA complexes protein-DNA cross-links are reversed and the DNA is extracted. Various methods exist to identify binding sites and determine relative occupancy of the protein of interest; these include quantitative PCR, probing microarrays or sequencing the isolated DNA (ChIP-seq). This chapter details the method of chromatin immunoprecipitation of TOP2 to the point of DNA extraction from the precipitated protein-DNA complexes. |
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