pSP73载体介绍:
The pSP73 Vector offers a wide range of restriction sites, providing greater versatility in cloning and transcription of RNA in vitro. The pSP73 Vector contains the SP6 and T7 RNA polymerase promoters and a unique multiple cloning region, which includes restriction sites for BglII, EcoRV, ClaI, EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, AccI, PstI, SphI, HindIII, PvuII and XhoI. The pSP72 and pSP73 Vectors are essentially identical except for the orientation of the multiple cloning region.
质粒类型:大肠杆菌表达载体
高拷贝/低拷贝:高拷贝
克隆方法:多克隆位点,限制性内切酶
载体大小:2464bp
载体抗性:Ampicillin (氨苄青霉素)
启动子:T7
复制子:ColE1 ori
质粒分类:大肠杆菌载体;克隆质粒
质粒大小:2464bp
原核抗性:氨苄青霉素Amp
克隆菌株:DH5α培养条件:37℃,有氧,LB
5'测序引物:Sp6:ATTTAGGTGACACTATAGAA
3'测序引物:T7:TAATACGACTCACTATAGGG
使用方法:
1.收到质粒后,5000rpm离心30s。
2.取2ul加入到100ul感受态细胞中,混匀;冰浴30min,42℃热击90s,再冰浴2min。
3.加入液体培养基,180rpm震荡培养。离心收集菌体,全部涂布至固体LB平板。
4.挑取平板上的单克隆菌落,接种至LB液体培养基37℃震荡培养12-16h。
5.提取质粒用于后续实验,暂时不用的质粒可以保存于-20℃。