Methylmercury as a reversible denaturing agent for agarose gel electrophoresis

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摘要：

A method for agarose gel electrophoresis under denaturing conditions, with methylmercuric hydroxide as the denaturing agent, is described. Methylmercuric hydroxide is a strong and reversible denaturing agent. It appears that complete denaturation of any base-paired secondary structural feature of a nucleic aicd can be achieved at practical concentrations of CH3HgOH. The method has been tested by showing that singly nicked circular duplex PM2 DNA is dissociated into more rapidly migrating linear and circular single-strand forms by a CH3HgOH concentration within 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 rRNA, the melting transition occurs mainly in 2–3 mm range of CH3HgOH. For a series of RNA's, 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, just as in other gel electrophoresis systems.

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Methylmercury Compounds DNA DNA, Circular RNA Electrophoresis Indicators and Reagents Nucleic Acid Denaturation Molecular Weight