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Formamide deionized

甲酰胺去离子

公司名称: AppliChem
产品编号: A2156
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KMnO4 Footprinting
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2012-11-05
[Abstract]  The KMnO4 footprinting method offers a rapid and easy way to detect and localize single-stranded regions within a duplex DNA molecule, such as it occurs for instance within an actively transcribing RNA polymerase-DNA complex or during R-loop formation in DNA-RNA hybrid structures. The method is based on the selective oxidation of single-stranded thymines in DNA. The modified nucleotides react with strong bases by ring opening and subsequent phosphodiester cleavage. Because the modified nucleotides will not be recognized by DNA polymerase sites of modification can also be analyzed ... [摘要]  KMnO 4足迹法提供了检测和定位双链DNA分子内的单链区域的快速和简单的方法,例如其在活性转录的RNA聚合酶-DNA复合物内或在R期间发生在DNA-RNA杂交结构中形成。该方法基于DNA中单链胸腺嘧啶的选择性氧化。修饰的核苷酸通过开环和随后的磷酸二酯切割与强碱反应。因为修饰的核苷酸不能被DNA聚合酶识别,还可以通过用Klenow DNA聚合酶引物延伸来分析修饰的位点,其在修饰前停止延伸一个残基。因此,修饰的碱基位置的定位可以在变性聚丙烯酰胺凝胶上在哌啶催化磷酸二酯切割3'-或5'-端32 P-端标记的DNA之后进行,或者通过用非 - 使用32 P标记的寡核苷酸引物。由于KMnO 4可以穿透膜的事实,足迹法也可以用于活细胞内的足迹分析。

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