一种经济、简便和准确鉴定重组质粒的方法

An Economic,Simple and Accurate Method for Identifing Recombinant Plasmids

目的:提高重组质粒的筛选效率,探讨一种经济、快速并准确的重组质粒鉴定方法。方法:取200μl菌液于Eppendorf管中高速离心1min,倒出上清,沉淀中加入20μl裂解缓冲液以裂解细胞并释放出内部质粒,10000r/min离心10min后取5～10μl上清液进行琼脂糖凝胶电泳,并比较其迁移位置来以判断重组质粒。结果:该方法鉴定结果和PCR鉴定结果完全一致。结论:该方法具有成本低廉、简便快速的特点,在对大量样品进行鉴定时具明显优势,适合在常规实验室推广。

Objective：To improve the efficiency of screening recombinant plasmids,an economic,rapid and accurate identification method was explored.Method：200μl bacterial cells have been centrifuged in the eppendorf tube at high-speed for 1min,cells were precipitated and the supernatants were removed.20μl lysis buffer was added to the precipitated to release the plasmid,after the solution was centrifuged at 10 000r/min for 10 minutes,5-10μl supernatants were analyzed by electrophoresis on agarose gel,the recombinant plasmids have been judged by comparison of their migration position.Result：The identified results using this method was the same to the results identified by PCR.Conclusion：The method has the characteristics of low-cost,simple and rapid,which has obvious advantages when a large number of samples to be identified,it is suitable to be spreaded in other routine laboratories.