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- 详细信息
- 文献和实验
- 技术资料
- 库存:
140
- 英文名:
pET-32a(+)-RNA-directed-RNA-polymerase plasmid
- 保质期:
2年
- 供应商:
上海烜雅生物科技有限公司
- 保存条件:
2-8℃
- 规格:
质粒干粉
英文名称:pET-32a(+)-RNA-directed-RNA-polymerase plasmid
形式:质粒干粉
参考用途:质粒载体
培养基:酵母膏:5.0,蛋白胨:10.0,NaCl:10.0,pH:7.0(25℃)
生长条件:LB+氨苄青霉素,37℃,克隆菌株BL21(DE3)
存储条件:2-8℃
安全等级:1
操作步骤:
1.溶解: 收到质粒干粉后,请加入 20μl 无菌水至管底,溶解质粒,室温静置 1min;
2.混合(吸附质粒): 200μl 感受态细胞 + 质粒 DNA 5~10µl 混匀,冰上放置 30min;
3.热激导入: 42℃静置 90s;
4.收缩膜孔: 冰浴 2min;
5.修复培养: 毎管加 800µl LB 液体培养基,37 ℃培养 1h 150 r/min;
6.筛选培养: 将适当体积(100 µl)的复苏细胞,涂布在相应抗性的 LB 平板上,正置平皿 30min(琼脂面务必干燥后), 倒置培养 12-16h,出现菌落。
7.提取: 挑取单克隆菌落至相应抗性 LB 液体培养基中,震荡培养 12-16h,根据试验需要提取质粒。
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