This is a detailed experimental protocol for the transformation of plasmid DNA into competent E. coli cells, specifically using the DH5α strain. The protocol encompasses the entire process, from the preparation of LB agar plates supplemented with ampicillin to the final inoculation of transformed cells in LB broth. Key steps include the careful thawing of competent cells, the addition of plasmid DNA, a precise heat shock treatment at 42°C for enhanced plasmid uptake, and subsequent recovery and plating of the cells. The methodology is designed to maximize the efficiency of transformation, crucial for subsequent molecular biology experiments such as gene cloning and expression analysis. This protocol serves as a fundamental guide for researchers in the field of molecular genetics and microbiology.