遗传 ›› 2011, Vol. 33 ›› Issue (4): 404-410.doi: 10.3724/SP.J.1005.2011.00404

• 技术与方法 • 上一篇    

4C-克隆筛选过程中的质量控制

田靖, 陈娜, 赵志虎, 陈惠鹏   

  1. 军事医学科学院生物工程研究所, 北京 100071
  • 收稿日期:2011-01-21 修回日期:2011-03-16 出版日期:2011-04-20 发布日期:2011-04-25
  • 通讯作者: 赵志虎;陈惠鹏 E-mail:zhaozh@nic.bmi.ac.cn;ChenHP0909@163.com
  • 基金资助:

    国家自然科学基金项目(编号:30871374)资助

The quality control of 4C-clone screening assay

TIAN Jing, CHEN Na, ZHAO Zhi-Hu, CHEN Hui-Peng   

  1. Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China
  • Received:2011-01-21 Revised:2011-03-16 Online:2011-04-20 Published:2011-04-25
  • Contact: ZHAO Zhi-Hu;CHEN Hui-Peng E-mail:zhaozh@nic.bmi.ac.cn;ChenHP0909@163.com

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: PCR 扩增, 染色体相互作用, 质量控制, SOX2, 环形染色体构象俘获

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.

Key words: circular chromosome conformation capture, chromosomal interaction, quality control, SOX2, PCR amplification