大肠杆菌DH5a菌株高效电击转化条件的优化
Fujian Agricultural Science and Technology（2023）
Abstract
为探索大肠杆菌DH5a高效电击转化条件,通过对电击转化条件中细胞生长状态、DNA加入量、感受态细胞体积、电场强度、速冻和恢复时间等关键因素进行优化,筛选出大肠杆菌DH5a高效电击转化条件.结果表明:大肠杆菌在细胞生长状态OD600值为0.6时制备感受态细胞,按每管50 μL体积分装、-80℃乙醇速冻后,加入10 pg pUC 19质粒DNA,在20 kv·cm-1电场强度、电容25 μF、电阻200 Ω条件下电击转化,转化后恢复培养60 min,转化效率达1010 cfu·μg-1 DNA以上水平,可满足基因文库构建、抗体库筛选等试验需求.
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