产品介绍
pCAMBIA1301-35SN植物表达质粒
基本信息
启动子: CaMV 35S
复制子: pVS1 oriV,ori
质粒分类: 植物系列,蛋白过表达载体
原核抗性: Kan
筛选标记: Hyg
克隆菌株: DH5α
培养条件: 37℃,有氧 LB
表达宿主: 植物细胞
5'测序引物: M13F:TGTAAAACGACGGCCAGT
3'测序引物: 根据序列设计引物
质粒简介
This vector contains a fully functional gusA reporter construct for simple and sensitive analysis of gene function or presence in regenerated plants by GUS assay. The construct uses E.coli gusA with an intron (from the castor bean catalase gene) inside the coding sequence to ensure that expression of glucuronidase activity is derived from eukaryotic cells, not from expression by residual A.tumefaciens cells. This vector is suitable for insertion of other genes of interest containing their own promoter and terminator. Researchers can excise the gusA gene and insert their own gene of interest in its place or use these vectors to create fusions of gusA with their gene of interest. These vectors contain the pUC18 polylinker-lacZa.
质粒图谱
pCAMBIA1301-35SN植物表达质粒使用说明:
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。
pCAMBIA1301-35SN植物表达质粒注意事项:
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。