Abstract
Chromatin immunoprecipitation (ChIP) of epigenetic modifications (epiChIP) coupled with quantitative PCR (qPCR), is the current Gold Standard in epigenetic research and will continue to be so for the foreseeable future. The combination of epiChIP and qPCR seems like a match made in heaven, but in reality, it is a relationship fraught with obstacles and hidden pitfalls and generating high quality ChIP data takes time, patience and practice. This chapter aims towards outlining the history and development of ChIP-qPCR, highlight inherent challenges and illuminate pathways around obstacles by discussing the key component parts of the assay: affinity reagents, chromatin preparation, dynamic range, chromatin precipitation, qPCR and data analysis and presentation. The aim is to outline the steps needed to successfully harness the full power of the ChIP/epiChIP assay to generate reliable and reproducible results.
| Original language | English (US) |
|---|---|
| Title of host publication | Epigenetics Methods |
| Publisher | Elsevier |
| Pages | 281-296 |
| Number of pages | 16 |
| ISBN (Electronic) | 9780128194140 |
| DOIs | |
| State | Published - Jan 1 2020 |
Keywords
- ChIP
- ChIP data analysis
- ChIP data presentation
- ChIP titration
- ChIP-qPCR
- Chromatin
- Chromatin size distribution
- Crosslinking
- MNAse
- N-ChIP
- NEXSON
- Sonication
- X-ChIP
- epiChIP
- qPCR
ASJC Scopus subject areas
- General Medicine
- General Biochemistry, Genetics and Molecular Biology