产品详情
pMDLg-pRRE慢病毒包装质粒
基本信息
启动子: CMV
复制子: pUC ori
终止子: β-globin poly(A)
质粒大小: 8890bp
原核抗性: Amp
克隆菌株: Stbl3
培养条件: 37℃,有氧 LB
表达宿主: 哺乳细胞
诱导方式: 无须诱导,瞬时表达
5'测序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: 根据序列设计引物
质粒简介
This 3rd generation LV packaging plasmid includes gag, coding for the virion main structural proteins; pol, responsible for the retrovirus-specific enzymes; and RRE, a binding site for the Rev protein which facilitates export of the RNA from the nucleus. This plasmid does not include Rev. The third generation packaging system offers maximal biosafety but is more cumbersome, as it involves the transfection of four different plasmids in the producer cells.
质粒图谱
pMDLg-pRRE慢病毒包装质粒使用说明:
1、收到质粒干粉后请先5000rpm离心1min,再加入20μl无菌水溶解质粒,室温放置1min;
2、从-80℃冰箱中取出相应的感受态,置于冰盒上解冻,并做好标记;
3、取2μl质粒加至100μl感受态中,冰浴30min;
4、42℃热激90s,再冰浴2min;
5、加入900μl无抗的LB液体培养基,180rpm震荡培养45min;
6、6000rpm离心5min,仅留100ul上清混匀菌体沉淀;
7、混匀后的菌液加至对应抗性的LB平板上,倒入适量玻璃珠,涂匀液体;
8、将平板正向培养1h,再倒置培养12h~16h;
9、挑取单克隆菌落至对应抗性的LB液体培养基中,震荡培养12h~16h,根据实验需要提取质粒。
pMDLg-pRRE慢病毒包装质粒注意事项:
1、如果您收到的是甘油菌种,请先四区划线,挑取单克隆培养。
2、如果第二天转化平板长的过多,请将质粒按比例稀释后再转化。