Chromatin Immunoprecipitation of Low Number of FACS-Purified Epidermal Cells.
Chromatin Immunoprecipitation of Low Number of FACS-Purified Epidermal Cells.
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DOI：
10.1007/978-1-0716-0648-3_17
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Ezhkova E
中科院分区：
文献类型：
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作者：
Bar C;Valdes VJ;Ezhkova E
染色质免疫沉淀测序(CHIP-SEQ)是一种检测染色质与其结合蛋白之间相互作用的方法。这种方法已经被广泛用于描述许多细胞类型的表观遗传景观，然而，一个限制因素是对大量细胞的要求。在这里，我们描述了一种用于新生小鼠表皮细胞芯片的方案，该芯片通过荧光激活细胞分选(FACS)纯化。该方法专门针对预先固定的低细胞数量进行了优化，从而产生了足够的免疫沉淀DNA，适合全基因组分析。
Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a method designed to detect interactions between chromatin and the proteins bound to it. This method has been widely used for characterizing epigenetic landscapes in many cell types, however, a limiting factor has been the requirement of a high number of cells. Here, we describe a protocol for ChIP in epidermal cells from a newborn mouse, purified by fluorescence- activated cell sorting (FACS). This protocol has been optimized specifically for pre-fixed, low cell numbers, resulting in enough immunoprecipitated DNA suitable for genome-wide analysis.