Construction of plasmids containing site-specific DNA interstrand cross-links for biochemical and cell biological studies.
Construction of plasmids containing site-specific DNA interstrand cross-links for biochemical and cell biological studies.
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DOI：
10.1007/978-1-61779-998-3_15
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发表时间：
2012
期刊：
影响因子：
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通讯作者：
Scharer, Orlando D
中科院分区：
文献类型：
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作者：
Enoiu, Milica;Ho, The Vinh;Long, David T;Walter, Johannes C;Scharer, Orlando D
含有位点特异性DNA链间交联(ICL)的质粒是在生物化学和细胞水平上研究ICL修复途径的宝贵工具。我们描述了一种在特定位置含有单个ICL的质粒DNA底物的制备方法。该程序是通用的，导致可靠的纯DNA底物的产生，并适合于将几乎任何类型的DNA损伤整合到质粒中。
Plasmids containing a site-specific DNA interstrand cross-link (ICL) are invaluable tools for the investigation of ICL repair pathways at the biochemical and cellular level. We describe a procedure for preparation of plasmid DNA substrates containing a single ICL at a specific site. The procedure is versatile, leads to reliable yields of pure DNA substrate, and is suitable for the incorporation of virtually any type of DNA lesion into plasmids.