Abstract
Chromatin immunoprecipitation (ChIP) combined with high-throughput sequencing (ChIP-seq) has become the gold standard for whole-genome mapping of protein-DNA interactions. However, conventional ChIP protocols necessitate the use of large numbers of cells, and library preparation steps associated with current high-throughput sequencing platforms require substantial amounts of DNA; both of these factors preclude the application of ChIP-seq technology to many biologically important but rare cell types. Here we describe a nano-ChIP-seq protocol that combines a high-sensitivity small-scale ChIP assay and a tailored procedure for generating high-throughput sequencing libraries from scarce amounts of ChIP DNA. In terms of the numbers of cells required, the method provides two to three orders of magnitude of improvement over the conventional ChIP-seq method and the entire procedure can be completed within 4 d.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1656-1668 |
| Number of pages | 13 |
| Journal | Nature Protocols |
| Volume | 6 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2011 |
Funding
acKnowleDGMents We acknowledge all the members of the Bernstein Lab for constructive discussions during the method development. We thank M. Coyne, S. Furuyama, S. Gillespie and N.A. Okan for their critical reading of the manuscript and J. Zhu, N. Shoresh and T. Mikkelsen for computational assistance. This research was supported by funds from the Starr Cancer Consortium, a Charles E. Culpeper Scholarship, the US National Human Genome Research Institute, the National Institutes of Health Roadmap for Epigenomics and the National Heart, Lung and Blood Institute.
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology