ChIP-Seq of chromatin marks at distal enhancers in Mouse Embryonic Stem Cells and adult tissues.. Mus musculus

Enhancers are enriched by histone H3 lysine 4 mono methylation (H3K4me1). To gain insight into the relation between enhancers and developmental state, chromatin immunoprecipitation coupled with massive parallel sequencing (ChIP-seq) was performed in several cell types to determine the genome-wide binding targets of H3K4me1 and several other possible enhancer associated modifications. Overall design: DNA was enriched by chromatin immunoprecipitation (ChIP) and analyzed by Solexa sequencing. A chromatin IP against histone H3 or whole cell extract was sequenced and used as the background to determine enrichment. ChIP was performed mainly using antibodies against the chromatin marks H3K4me1 and H3K27ac.

增强子（enhancers）富集于组蛋白H3赖氨酸4单甲基化（H3K4me1）修饰。为深入解析增强子与发育状态之间的关联，研究人员在多种细胞类型中开展了染色质免疫共沉淀结合高通量测序（ChIP-seq）实验，以确定全基因组范围内H3K4me1以及其他若干潜在增强子相关组蛋白修饰的结合靶标。总体实验设计如下：通过染色质免疫共沉淀（ChIP）富集DNA片段，并采用索莱拉（Solexa）测序进行分析。以针对组蛋白H3的染色质免疫沉淀产物及全细胞提取物的测序结果作为背景对照，用于判定富集程度。本次ChIP实验主要使用靶向染色质修饰H3K4me1与H3K27ac的抗体完成。