Plasmid DNA isolation using alkaline lysis method from E. coli DH5α cells

This chapter focuses on the alkaline lysis method for plasmid DNA isolation, which is probably one of the generally used methods for the isolation of circular DNA from bacterial cells. It enables rapid annealing following denaturation of plasmid DNA, which causes its separation from the bacterial chromosome without the activation of the enzymes responsible for DNA digestion. The alkaline environment of the solution containing bacterial DNA helps in faster renaturation of plasmid DNA than genomic DNA. The DNA obtained using this technique is highly purified. Specific protocols for alkaline lysis vary from lab to lab.