Maximizing Genetic Transformation Success with Specialized Bacterial Cells

Electrocompetent cells refer to bacteria that have been chemically or physically treated to make them competent for transformation via electroporation. Electroporation is a process where an electric pulse is used to create temporary pores in the bacterial cell membrane through which exterior DNA can enter.

Preparing Bacterial Cells for Electroporation
In order to render bacterial cells electrocompetent, chemical treatments are used to weaken the cell wall and membrane. This weakening allows pores to form in the presence of an electric field. Common electrocompetent cell preparation protocols involve treating cells with solutions like calcium chloride, instead of potassium chloride or rubidium chloride. These divalent cations displace magnesium ions from the cell surface, disrupting the integrity of the outer membrane and cell wall.

Cells are also chilled on ice during preparation to further weaken the membrane. Washing steps help remove charged particles and metabolites that could inhibit transformation. Dilution of the cell suspension increases electrocompetency by lowering cell density and limiting cellular interactions during electroporation. Properly prepared Electrocompetent Cells are then ready for genetic manipulation through electroporation.

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