摘要
目的介绍一种快速获取高纯度质粒DNA的方法。方法使用替代试剂,建立改良的碱裂解法进行质粒DNA提取,并用琼脂糖电泳验证所得的质粒的质量,紫外测量估算产量及纯度。限制性酶切验证其用度。结果琼脂糖电泳显示质粒DNA条带明亮,以超螺旋方式为主。A260/280为1.903,含量为320μg/mL,酶切后得到约500bp的目的片段。结论改良碱裂解法操作简单、实用,所得样品纯度高,达到了分子生物学常规实验的要求。
[Objective] Introduce a method for rapid extraction of Superior-Quality Plasmid DNA. [Method] Use alternative reagent to establish modified alkaline lysis method for extraction of plasmid DNA and use agarose eleetrophoresis to examine the quality of plasmid DNA, ultraviolet speetrophotometry to evaluate its production and purity, restricted enzyme cutting to examine its availability. [Results] The agarose eleetrophoresis showed that the plasmid DNA band was bright, mainly by form of superhelix. A260/280 was 1.903, DNA concentration was 320μg/ ml, and we got 500 bp target fragment by restricted enzyme cutting. [Conclusion] We can get high purity of sample DNA by this simple and practical new method, it can meet the requirement of routine molecular biochemistry experiments.
出处
《中国医学工程》
2006年第2期199-201,共3页
China Medical Engineering
关键词
质粒
纯化
碱裂解法
plasmid
purifing
alkaline lysis
