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文献和实验Transformation of E. coli by Electroporation
Materials: SOB medium E. coli host strain such as DH5α WB tRNA 5 M ammonium acetate
In recent years several techniques have been described for the introduction of DNA molecules into Escherichia coli . These are based on the findings of Mandel and Higa (1 ), who demonstrated that incubation of cells with naked bacteriophage DNA
1.O/N (5ml E.coli in LB-->1ml into 200ml LB/37℃ 2.Grow 2-3H to A660 ~0.3-0.4 3.Chill cells 10'/ice Spin down 10',5K,4℃ 4.Wash 1X 200ml 10% glycerol (sterile)40℃; Spin down 10',5K,4℃ 5.Wash 1X 40 ml 10% glycerol--> transfer to S34 tubes & spin
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