Sokolova Maria, Vartiainen Maria K
Institute of Biotechnology, University of Helsinki, Helsinki, Finland.
Methods Mol Biol. 2023;2626:335-351. doi: 10.1007/978-1-0716-2970-3_18.
Chromatin is composed of DNA and its associated proteins, and has an essential role in all cellular processes, including those taking place during Drosophila oogenesis. In order to understand the molecular basis of chromatin-based processes, such as transcription, it is essential to be able to study how and when different proteins, such as transcription factors, histones and RNA polymerases, interact with chromatin. One of the most popular methods to study this is chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq). Here, we describe a ChIP-seq protocol that has been optimized for Drosophila ovaries, focusing on sample preparation through preliminary data processing.
染色质由DNA及其相关蛋白质组成,在所有细胞过程中都起着至关重要的作用,包括果蝇卵子发生过程中发生的那些过程。为了理解基于染色质的过程(如转录)的分子基础,能够研究不同蛋白质(如转录因子、组蛋白和RNA聚合酶)如何以及何时与染色质相互作用至关重要。研究这一问题最常用的方法之一是染色质免疫沉淀测序(ChIP-seq)。在此,我们描述了一种针对果蝇卵巢优化的ChIP-seq方案,重点是从样本制备到初步数据处理。