METHYLMERCURY AS A REVERSIBLE DENATURING AGENT FOR AGAROSE-GEL ELECTROPHORESIS
METHYLMERCURY AS A REVERSIBLE DENATURING AGENT FOR AGAROSE-GEL ELECTROPHORESIS
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DOI：
10.1016/s0003-2697(76)80049-8
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发表时间：
1976-01-01
影响因子：
2.9
通讯作者：
DAVIDSON, N
中科院分区：
文献类型：
--
作者：
BAILEY, JM;DAVIDSON, N
本文介绍了一种以氢氧化甲基汞（CH_3 HgOH）为变性剂的变性琼脂糖凝胶电泳方法。CH 3 HgOH是一种强的可逆变性剂。核酸的任何碱基配对二级结构特征的完全变性显然可以在CH 3 HgOH的实际浓度下实现。该方法的测试结果表明，当CH 3 HgOH浓度在2.5-5.0 mM范围内时，单切口环状双链PM2病毒DNA会解离为更快速迁移的线性和环状单链形式。在足够的CH 3 HgOH存在下，单链RNA分子的迁移率降低，因为它们的二级结构被破坏。对于HeLa 28 S核糖体RNA，解链转变主要发生在2-3 mM范围的CH 3 HgOH中。对于一系列RNA（大肠杆菌16和23 S RNA），在5 mM CH 3 HgOH（对应于完全变性）下，log（分子量）与电泳迁移率呈线性相关，log（迁移率）与琼脂糖浓度呈线性相关，与其他凝胶电泳系统相同。
A method for agarose gel electrophoresis under denaturing conditions, with methylmercuric hydroxide (CH3HgOH) as the denaturing agent, is described. CH3HgOH is a strong and reversible denaturing agent. Complete denaturation of any base-paired secondary structural feature of a nucleic acid can apparently be achieved at practical concentrations of CH3HgOH. The method was tested by showing that singly nicked circular duplex PM2 virus DNA is dissociated into more rapidly migrating linear and circular single-strand forms by a CH3HgOH concentration in the 2.5-5.0 mM range. The mobilities of single-strand RNA molecules are decreased in the presence of sufficient CH3HgOH because their secondary structure is disrupted. For HeLa 28S ribosomal RNA, the melting transition occurs mainly in 2-3 mM range of CH3HgOH. For a series of RNA''s (Escherichia coli 16 and 23 S RNA), at 5 mM CH3HgOH, corresponding to complete denaturation, there is a linear dependence of log (molecular weight) on electrophoretic mobility and of log (mobility) on agarose concentration, as in other gel electrophoresis systems.