Chemical transformation of E. Coli cells is the process by which bacterial cell asre made to take up desired plasmids. Chemical transformation is performed after TOPO cloning. Some preparatory steps include equilibrating the water bath to 420C, warming selective plates at 720C for 30 minutes and thawing vials of One Shot Top 10 E.Coli cells on ice. To start off, 2 microliters of TOPO cloning reaction products are added into vials, mixed gently and then incubated on ice for 5-30 minutes. The cells are heat shocked for 30 second at 420C without shaking before being placed on ice for 2 minutes. 250 microliters of S.O.C. medium is added, the tube is capped tightly and then shook horizontally at 200RPM and 370C for 1 hour.
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