Title
Analysis of long-range chromatin interactions using Chromosome Conformation Capture
UMMS Affiliation
Program in Systems Biology; Program in Gene Function and Expression; Department of Biochemistry and Molecular Pharmacology
Publication Date
2012-11
Document Type
Article
Subjects
Chromatin; Chromosomes; Nucleic Acid Conformation
Disciplines
Genetics and Genomics | Systems Biology
Abstract
Chromosome Conformation Capture, or 3C, is a pioneering method for investigating the three-dimensional structure of chromatin. 3C is used to analyze long-range looping interactions between any pair of selected genomic loci. Most 3C studies focus on defined genomic regions of interest that can be up to several hundred Kb in size. The method has become widely adopted and has been modified to increase throughput to allow unbiased genome-wide analysis. These large-scale adaptations are presented in other articles in this issue of Methods. Here we describe the 3C procedure in detail, including the appropriate use of the technology, the experimental set-up, an optimized protocol and troubleshooting guide, and considerations for data analysis. The protocol described here contains previously unpublished improvements, which save time and reduce labor. We pay special attention to primer design, appropriate controls and data analysis. We include notes and discussion based on our extensive experience to help researchers understand the principles of 3C-based techniques and to avoid common pitfalls and mistakes. This paper represents a complete resource and detailed guide for anyone who desires to perform 3C.
DOI of Published Version
10.1016/j.ymeth.2012.07.022
Source
Methods. 2012 Nov;58(3):192-203. doi: 10.1016/j.ymeth.2012.07.022. Epub 2012 Aug 15. Link to article on publisher's site
Journal/Book/Conference Title
Methods (San Diego, Calif.)
Related Resources
PubMed ID
22903059
Repository Citation
Naumova, Natalia; Smith, Emily M.; Zhan, Ye; and Dekker, Job, "Analysis of long-range chromatin interactions using Chromosome Conformation Capture" (2012). Program in Systems Biology Publications and Presentations. 18.
https://escholarship.umassmed.edu/sysbio_pubs/18
Comments
Co-author Emily Smith is a doctoral student in the Interdisciplinary Graduate Program in the Graduate School of Biomedical Sciences (GSBS) at UMass Medical School.