质粒DNA提取方法的探讨

本实验利用构建好的含有抗病虫基因的大肠杆菌的质粒,采用了碱裂解法、煮沸法、SDS法进行提取。在比较这些方法的同时,进一步对它们进行优化,从而使其得到完善。结果表明:在小质粒中,三种方法中碱裂解法所得到的DNA制品其浓度和纯度都明显高于其它两种方法。

A Research for Methods of Extracting Plasmid DNA

In laboratory utilized coliform plasmids,they can bring anti-sick and anti-insect gene into host cells to replicate and express as vectors,then the host cells display the character of goal gene controlling.In order to extract high qualitied and purified plasmids,we had used three methods: Alkaline-lysis method,boiling method,SDS method.In the contrast with them, we have improved them in order to perfect them.Results showed that: to small plasmids Alkaline-lysis method of them obtain DNA plasmids,its quality and purify are higher than other methods'.SDS method is mild and fit for over 15Kb plasmids.Boiling operating is simple,but it can't get rid of protein completely and its concentration and purify are obviously lower than other methods.