pGL4.22[luc2CPPuro]哺乳报告质粒

保存条件 ：负20度

保质期 ：3个月

库存 ：100

供应商 ：钦胜生物

规格 ：2UG

产品介绍

pGL4.22[luc2CP/Puro]哺乳单荧光素酶报告质粒

 

基本信息

 

复制子: pUC

原核抗性: 氨苄青霉素Amp

筛选标记: 嘌呤霉素Puro

克隆菌株: 大肠杆菌DH5α

培养条件: 37℃

 

质粒简介

             pGL4.22[luc2CP/Puro]是一个哺乳细胞单荧光素酶报告质粒。The pGL4.22[luc2CP/Puro] Vector encodes the luciferase reporter gene luc2CP (Photinus pyralis) and is designed for high expression and reduced anomalous transcription. This vector contains a mammalian selectable marker for puromycin resistance in which the number of transcription factor binding sites has been reduced and mammalian codon usage optimized. The vector is engineered with fewer consensus regulatory sequences for reduced background and a decreased risk of anomolous transcription and has a synthetic reporter gene, which has been codon optimized for mammalian expression. The pGL4.22[luc2CP/Puro] Vector is a basic vector with no promoter. However, it contains a multiple cloning region that allows cloning of a promoter of choice. The luc2CP reporter gene contains two protein destabilization sequences, hCL1 and hPEST. The protein encoded by luc2CP responds more quickly and with greater magnitude to changes in transcriptional activity than the luc2 gene, its more stable counterpart.

 

质粒图谱

 

 

pGL4.22[luc2CP/Puro]哺乳单荧光素酶报告质粒使用说明：

    

     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。

 

pGL4.22[luc2CP/Puro]哺乳单荧光素酶报告质粒注意事项：

 

      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。