Inoue H, Nojima H, Okayama H
Research Department, Turuga Enzyme Plant, Toyobo Co, Ltd., Fukui-ken, Japan.
Gene. 1990 Nov 30;96(1):23-8. doi: 10.1016/0378-1119(90)90336-p.
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 x 10(9) cfu/microgram 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 particularly useful for construction of high-complexity cDNA libraries with a minimum expenditure of mRNA.
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）。用SEM制备的细胞（DH5、JM109和HB101）具有极高的转化能力（1 - 3×10⁹ cfu/微克pBR322 DNA），并且可以在液氮中保存至少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转化时，DNA制备过程中的盐对其影响极小。这些感受态细胞对于以最少的mRNA用量构建高复杂性cDNA文库特别有用。
