Rapid Isolation and Purification of Plasmid DNA Using High Performance Liquid Chromatography

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High Performance Liquid Chromatography (HPLC) has been employed as an analytical tool for the purification and separation of nucleic acids. A Nucleogen DEAE 4000-10 weak anion exchange column, prepacked with modified silica gels, was used to purify and separate a number of Escherichia coli plasmids. Plasmid DNAs were extracted by the alkaline lysis method. The cleared lysate was injected directly onto the Nucleogen column, and the peaks were collected, desalted and analysed by gel electrophoresis. On the chromatogram, the pBR322 formed a distinctive peak at 27 minutes and partial separation was made for the E. coli V517 plasmids. Plasmid pBR322 ... continued below

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v, 48 leaves : ill.

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Nam, Kiebang May 1987.

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  • Nam, Kiebang

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High Performance Liquid Chromatography (HPLC) has been employed as an analytical tool for the purification and separation of nucleic acids. A Nucleogen DEAE 4000-10 weak anion exchange column, prepacked with modified silica gels, was used to purify and separate a number of Escherichia coli plasmids. Plasmid DNAs were extracted by the alkaline lysis method. The cleared lysate was injected directly onto the Nucleogen column, and the peaks were collected, desalted and analysed by gel electrophoresis. On the chromatogram, the pBR322 formed a distinctive peak at 27 minutes and partial separation was made for the E. coli V517 plasmids. Plasmid pBR322 showed a clear band without any detectable contamination on agarose gel. This purified plasmid DNA is biologically active for enzymatic reaction commonly used in genetic engineering techniques.

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v, 48 leaves : ill.

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  • May 1987

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  • May 10, 2015, 6:16 a.m.

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  • Nov. 3, 2016, 8:31 a.m.

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Nam, Kiebang. Rapid Isolation and Purification of Plasmid DNA Using High Performance Liquid Chromatography, thesis, May 1987; Denton, Texas. (digital.library.unt.edu/ark:/67531/metadc504136/: accessed October 21, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; .