摘要
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.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第3期54-56,共3页
Biotechnology
基金
四川省教育厅基金项目(08Zb051)
乐山师范学院科研项目(Z0858)资助
关键词
重组质粒
裂解缓冲液
琼脂糖凝胶
电泳
recombinant plasmids
lysis buffer
agrose gel
electrophoresis