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Competent Cell Production
Competent cell production is the generation of bacterial cells into which exogenous DNA, such as cloned plasmids, can be introduced. Host cells for molecular cloning must be competent. Though host cells can be naturally competent, their competency is highly regulated. Therefore, host cells are made artificially competent by weakening their cell membrane or wall with chemicals. The most common method involves the repeated incubation of bacterial cells, such as E. coli, with cold calcium chloride. Calcium ions are believed to disrupt the negative charge of the cell wall lipopolysaccharides allowing the negatively charged DNA to enter the cell. Competent cells are weakened so care should be taken to maintain their viability. This includes storing them at -80 degrees C, thawing on ice, and avoiding excessive mechanical stress like pipetting. (Credit: Brooke Anderson-White)
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