4C-克隆筛选过程中的质量控制.pdfVIP

HEREDITAS (Beijing) 2011 年 4 月, 33(4): 1 ―7 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2011.00000 4C-克隆筛选过程中的质量控制 田靖, 陈娜, 赵志虎, 陈惠鹏 , 100071 摘要: 环形染色体构象俘获(Circular chromosome conformation capture, 4C)是一种高通量研究细胞染色体相互 作用、空间构象的技术。文章通过模拟的4C 样品, 优化反向PCR 条件, 建立了高效、特异的扩增方法, 对4C 克隆筛选等后续步骤进行严格的质量控制并对该方法的可行性进行了实例验证。该 4C-克隆筛选方法作为 4C 方法的质量控制标准, 具有重要的指导和监测作用。 关键词: 环形染色体构象俘获; 染色体相互作用; 质量控制; SOX2; PCR 扩增 The quality control of 4C-clone screening assay TIAN Jing, CHEN Na, ZHAO Zhi-Hu, CHEN Hui-Peng Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China Abstract: Circular chromosome conformation capture (4C) can be used to analyze the high-resolution interaction map of cis-regulatory elements in genome-wide studies. In this study, we optimized the condition of PCR with the mimical 4C sample in order to establish a specific and efficient assay, which was validated practically. This 4C-clone screening assay can be used as the quality control in the application of 4C assay. Keywords: circular chromosome conformation capture; chromosomal interaction; quality control; SOX2; PCR amplification , , , , , CT (Chromosome conforma- (Chromosome territory), tion capture, 3C) , [4] [1] , 3C , , DNA , [2] , , [3] (Circular chromosome conformation capture, 4C) 3C 收稿日期: 2011−01−21; 修回日