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《China Biotechnology》 2007-02
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Modification of Alkaline Lysis Method for Preparation of Plasmid DNAs from Recombinant E.coli

ZHANG Quan1 ZHU Hong-fei2 YU Ke-xiang1 XUE Fang-ming1 SUN Huai-chang1(1 Collge of Veteriniary Medicine Yangzhou University,Yangzhou 225009,China)(2 Institute of Animal Science and Veterinary Medicine Chinese Academy of Agricultural Science,Beijing 100081,China)  
In order to decrease the cost of the plasmid production,the standard alkaline lysis method for preparation of plasmid DNAs was modified,including the use of optimized volume of solution I to resuspend the centrifuged bacteria and a new solution III which contain less KAc to neutralize the cell lysate.Purification and quantitative analysis by BCA detection,Real-time PCR and agrose gel electrophoresis showed that both yield and quality of plasmid DNA prepared by the modified alkaline lysis remain unchanged compared to that by standard alkaline lysis method,but the cost was decreased to about 1/4 of normal method,which laid a solid foundation for large-scale production of recombinant plasmid DNAs for animal use.
【Fund】: 国家“863”计划资助项目(2005AA246020)
【CateGory Index】: Q78
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【Secondary Citations】
Chinese Journal Full-text Database 9 Hits
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