HIGH-EFFICIENCY TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS
HIGH-EFFICIENCY TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS
复制标题
DOI：
10.1016/0378-1119(90)90336-p
复制
发表时间：
1990-11-30
期刊：
影响因子：
3.5
通讯作者：
OKAYAMA, H
中科院分区：
文献类型：
--
作者：
INOUE, H;NOJIMA, H;OKAYAMA, H
我们重新评估了制备感受态大肠杆菌细胞的条件，并建立了一种简单高效的质粒转染方法（SEM）。通过SEM制备的细胞（DH5、JM109和HB101）具有极强的转化能力（每微克pBR322 DNA可产生1 - 3×10⁹个菌落形成单位），并且可以在液氮中储存至少40天而不丧失感受态。与电穿孔不同，使用这些感受态细胞进行的转化受DNA制备过程中盐分的影响极小。这些感受态细胞对于以最少的mRNA用量构建高复杂性的cDNA文库特别有用。
We have re-evaluated the conditions for preparing competent Escherichia coli cells and established a simple and efficient method (SEM) for plasmid transfection. Cells (DH5, JM109 and HB101) prepared by SEM are extremely competent for transformation (1-3 .times. 109 cfu/.mu.g of pBR322 DNA), and can be stored in liquid nitrogen for at least 40 days without loss of competence. Unlike electroporation, transformation using these competent cells is affected minimally by salts in DNA preparation. These competent cells are particulary useful for construction of high-complexity cDNA libraries with a minimum expenditure of mRNA.