UNIT 21.15 Chromatin Interaction Analysis Using Paired-End Tag Sequencing
Published Online: 1 JAN 2010
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Fullwood, M. J., Han, Y., Wei, C.-L., Ruan, X. and Ruan, Y. 2010. Chromatin Interaction Analysis Using Paired-End Tag Sequencing. Current Protocols in Molecular Biology. 89:21.15:21.15.1–21.15.25.
- Published Online: 1 JAN 2010
- Published Print: JAN 2010
Chromatin Interaction Analysis using Paired-End Tag sequencing (ChIA-PET) is a technique developed for large-scale, de novo analysis of higher-order chromatin structures. Cells are treated with formaldehyde to cross-link chromatin interactions, DNA segments bound by protein factors are enriched by chromatin immunoprecipitation, and interacting DNA fragments are then captured by proximity ligation. The Paired-End Tag (PET) strategy is applied to the construction of ChIA-PET libraries, which are sequenced by high-throughput next-generation sequencing technologies. Finally, raw PET sequences are subjected to bioinformatics analysis, resulting in a genome-wide map of binding sites and chromatin interactions mediated by the protein factor under study. This unit describes ChIA-PET for genome-wide analysis of chromatin interactions in mammalian cells, with the application of Roche/454 and Illumina sequencing technologies. Curr. Protoc. Mol. Biol. 89:21.15.1-21.15.25. © 2010 by John Wiley & Sons, Inc.
- DNA sequencing;
- 454 sequencing;
- Illumina sequencing;
- transcription factor binding sites;
- chromatin interactions;
- chromosome conformation capture;
- chromatin immunoprecipitation