摘要
在过去的十几年中,染色质免疫共沉淀测序技术(ChIP-seq)成为了分析表观基因组和鉴定DNA相关蛋白重要结合位点的主要技术。然而,ChIP-seq技术目前仍存在一些技术难点。其中一个重要限制是ChIP-seq需要大量的起始材料,需要至少数百万个细胞才能启动,然而实验中有两个关键步骤往往会造成所使用的样本丢失,即染色质制备和免疫沉淀。本实验通过从染色体制备入手,通过选择两种不同核酸水解酶DNase和MNase,设计不同实验组验证最佳酶切浓度,以优化ChIP-seq体系。结果证明0.1U/μL的MNase为本实验中ChIP-seq最佳酶切浓度。结果为ChIP-seq体系的优化提供了参考,对ChIP-seq技术在小鼠早期胚胎细胞中的应用以及发展有一定的积极意义。
In the past decade,Chromatin immunoprecipitation followed by sequencing,ChIP-Seq has become the main technology to analyze epigenome and identify important binding sites of DNA related proteins.However,chip-seq still has some technical difficulties.One important limitation is that chip-seq requires a large amount of starting material,which requires at least millions of cells to start.However,chromatin preparation and immunoprecipitation as two key steps in the experiment usually result in the loss of samples.This experiment started with chromosome preparation,in order to optimize the ChIP-seq system,two different nuclease hydrolases DNase and MNase were selected to design different experimental groups.The results showed that 0.1U/μL MNase was the optimal concentration of ChIP-seq.The results provide a reference for the optimization of ChIP-seq system,and have positive significance for the application and development of chip-seq technology in early mouse embryonic cells.
作者
高晗
钟蓓
Gao Han;Zhong Bei(College of Life Science Northeast Agriculture,Harbin Heilongjiang 150000,China)
出处
《现代畜牧科技》
2020年第1期8-11,共4页
Modern Animal Husbandry Science & Technology
