一种高效安全提取质粒DNA方法的建立  

Establishing an efficient and safe method of plasmid dna extraction

作　　者：张凤琴[1] 汤斌[1] 丁丽霞[1] 潘海波[1] 陈涛[1] 

机构地区：[1]安徽工程大学微生物发酵安徽省工程技术研究中心,安徽芜湖241000

出　　处：《安徽工程大学学报》2011年第2期4-6,共3页Journal of Anhui Polytechnic University

基　　金：安徽省高校自然科学研究重点基金资助项目(kj2007a018)

摘　　要：在碱裂解法提取质粒DNA的基础上做了改进,将NaOH浓度从0.20mol/L降低为0.10mol/L,使得超螺旋DNA的得率进一步提高.采用silica法纯化质粒DNA,避免了使用有毒性和挥发性气味的饱和酚和氯仿,使得操作更安全简单.电泳结果表明,提取的质粒DNA质量好,得率高,并且可以直接进行下一步的酶切、连接及转化等实验.同时,该方法中使用的硅胶可以回收再生进行重复利用,节省了实验费用.A new,safe and efficient plasmid extraction method modified on the basis of alkaline lysis method was established,which changed alkaline lysis solutionⅡformula with the reduction of NaOH concentration from 0.20 mol/L to 0.1 mol/L to increase the quantity of extracted superhelical DNA.In order to prevent the high toxicity and volatile smell produced by phenol and chloroform in alkaline lysis method,the silica was used in this method to purify plasmid DNA.The results showed that the quality of extracted plasmid DNA isolated by the established method was better than that of classical alkaline lysis method evaluated by the electrophoresis and spectrophotometer quantitative analytical methods.The restriction enzyme digestion experiment also proved that the plasmid produced by new method can be used effectively for further recombination manipulation such as ligation and transformation.Furthermore, silica used in this method can be recycled to,which reduces experimental expenses to a large extent.

关 键 词：碱裂解法 质粒DNA 超螺旋DNA silica法 

分 类 号：Q78[生物学—分子生物学]