The Mechanism of Transformation with Competent Cells

Learn that transformation is the process of introducing DNA into bacteria, which then replicate and express that DNA. Competent cells are bacteria capable of taking up exogenous DNA through artificial methods.

Use chemically competent cells treated with divalent cations to increase cell wall permeability. Apply a heat shock step to further improve DNA uptake into the bacterial cells.

Use an electrical field applied to competent cells in suspension to create temporary holes in the cell perimeter. This allows exogenous DNA to pass directly into the bacterial cytoplasm.

Grow the newly transformed bacteria in liquid medium without antibiotics to allow cellular repair and recovery. This outgrowth phase enables cells to stabilize after the transformation process.

Spread the recovered culture onto selective plates containing antibiotics to isolate only the transformant bacteria. Only cells that successfully acquired the DNA with antibiotic resistance genes will survive and grow.