Of a modified plasmid DNA extraction method for the establishment and application of.docVIP

 PAGE \* MERGEFORMAT 11 Of a modified plasmid DNA extraction method for the establishment and application of Authors: Zhang Linlin, Li Lei, Xiang Li, Da-Lin, Guo-Dong Zhu, WANG Xin-yang [Keywords:] plasmid DNA; extraction; digestion; transfection Abstract: Objective To establish a stable, reliable and improved method of extracting plasmid DNA and explore its experimental study in molecular biology in the application. Methods The purpose plasmid containing the amplified strains, using the improved alkaline lysis method a small amount of plasmid DNA extraction, trace extracts nucleic acid to measure the yield and purity of plasmid DNA; using agarose gel electrophoresis, endonuclease digestion and plasmid transfection of eukaryotic cells identified the quality and extracted plasmid DNA transfection efficiency. The results of modified plasmid DNA extraction method to obtain plasmid yield 3.2mg / L bacilli, A260/280 = 1.91; endonuclease digestion completely; plasmid DNA in order to super-helix structure of the main; eukaryotic cell transfection efficiency was about 20% . Conclusions Improved plasmid DNA extraction method is simple, practical, to obtain plasmid DNA production of more high quality, can achieve routine molecular biology experiments. Keywords: Plasmid DNA; extraction; digestion; transfection An improved and economical method for isolation of plasmid DNA ABSTRACT: Objective To explore a simple, improved and inexpensive method for isolation of plasmid DNA and its value in molecular biological laboratory research. Methods Plasmid DNA was isolated using general alkaline lysis method and improved method, respectively. The isolated plasmid DNA was quantified by spectrophotometric measurement, and evaluated by the electrophoresis of plasmid DNA in an agarose gel, the restriction enzyme digestion, and the transfection into eukaryotic cells. Results The quantity of plasmid DNA isolated by the improved method was 3.2μ g per milliliter culture of transform