产品详情
DH5αλpir大肠克隆菌(有kan,做结合转移实验未成功)
基本信息
产品名称:DH5αλpir大肠杆菌。E. coli DH5α λpir strain contains the pir gene (lysogenized lpir phage lysogen) and is designed for cloning and propagation of plasmids with R6K origin of replication sequence (like pUTmini-Tn5 vectors). To transfer this plasmids directly from DH5α λpir donor strain to the recipient strain it is necessary to perform a triparental mating with a helper strain such as E. coli DH5α (pRK2013) (see Helper E. coli strains). The genotype of the strain is sup E44, ΔlacU169 (ΦlacZΔM15), recA1, endA1, hsdR17, thi-1, gyrA96, relA1, λpir phage lysogen.
质粒图谱
DH5αλpir大肠克隆菌(有kan,做结合转移实验未成功)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。
DH5αλpir大肠克隆菌(有kan,做结合转移实验未成功)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。