产品介绍
pTagBFP-N哺乳荧光质粒
基本信息
启动子: CMV
复制子: pUC ori,f1 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳细胞,荧光蛋白报告载体
质粒大小: 4715bp
原核抗性: Kan
筛选标记: Neo
克隆菌株: DH5α
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: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
质粒简介
pTagBFP-N is a mammalian expression vector encoding blue fluorescent protein TagBFP. The vector allows generation of fusions to the TagBFP N-terminus and expression of TagBFP fusions or TagBFP alone in eukaryotic (mammalian) cells.
TagBFP codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. To increase mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the TagBFP coding sequence [Kozak, 1987]. Multiple cloning site (MCS) is located between PCMV IE and TagBFP coding sequence
质粒图谱
pTagBFP-N哺乳荧光质粒使用说明:
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。
pTagBFP-N哺乳荧光质粒注意事项:
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。