Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Nat Protoc. 2011 Sep 29;6(10):1656-68. doi: 10.1038/nprot.2011.402.
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.
染色质免疫沉淀（ChIP）联合高通量测序（ChIP-seq）已成为全基因组范围内蛋白质-DNA 相互作用图谱绘制的金标准。然而，传统的 ChIP 方案需要使用大量的细胞，而与当前高通量测序平台相关的文库制备步骤需要大量的 DNA；这两个因素都使得 ChIP-seq 技术无法应用于许多生物学上重要但数量稀少的细胞类型。在这里，我们描述了一种纳米 ChIP-seq 方案，该方案结合了高灵敏度的小规模 ChIP 检测以及一种针对从少量 ChIP DNA 生成高通量测序文库的定制程序。就所需的细胞数量而言，该方法比传统的 ChIP-seq 方法提高了两到三个数量级，整个过程可以在 4 天内完成。
