产品介绍
pET303/CT-His大肠表达质粒
基本信息
别名: pET303-CT-His
启动子: T7/lac promoter
复制子: ColE1 ori,F1 ori
终止子: T7 terminator
质粒分类: 大肠杆菌载体;PET系列表达质粒
质粒大小: 5369bp
质粒标签: C-6×His
原核抗性: 氨苄青霉素Amp
克隆菌株: 大肠杆菌DH5α
培养条件: 37℃,有氧,LB
表达宿主: 大肠杆菌BL21(DE3)
培养条件: 37℃,有氧,LB
诱导方式: IPTG或乳糖及其类似物
5'测序引物: T7:TAATACGACTCACTATAGGG
3'测序引物: T7-ter:TGCTAGTTATTGCTCAGCGG
质粒简介
pET303/CT-His pET303-CT-His是一个大肠杆菌pET系列表达质粒。The Champion pET302/NT-His and pET303/CT-His Vector Kit is designed for cloning a gene of interest via restriction enzyme(s) and ligase (REaL) and subsequent high-level expression from the strong bacteriophage T7 promoter. In addition to the T7 promoter, each vector contains only the necessary functional elements and an N- or C-terminal 6xHis tag (pET302/NT-His and pET303/CT-His, respectively) for convenient purification and detection (Figure 1). Expression levels obtained from these vectors may be higher than those obtained from another suppliers pET vector (Figure 2). To maximize expression, use with MagicMedia E. coli Expression Medium.
质粒图谱
pET303/CT-His大肠表达质粒使用说明:
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。
pET303/CT-His大肠表达质粒注意事项:
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。