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Transformation

Transformation is the process by which an organism acquires exogenous DNA. Transformation can occur in two ways: natural transformation and artificial transformation. Natural transformation describes the uptake and incorporation of naked DNA from the cell’s natural environment. Artificial transformation encompasses a wide array of methods for inducing uptake of exogenous DNA. In cloning protocols, artificial transformation is used to introduce recombinant DNA into host bacteria (E. coli). The most common method of artificial transformation of bacteria involves use of divalent cations (e.g., calcium chloride) to increase the permeability of the bacterium’s membrane, making them chemically competent, thereby increasing the likelihood of DNA acquisition. Another artificial method of transformation is electroporation, in which cells are shocked with an electric current, to create holes in the bacterial membrane. With a newly-compromised cell membrane, the transforming DNA is free to pass into the cytosol of the bacterium. Regardless of which method of tranformation is used, outgrowth of bacteria following transformation allows repair of the bacterial surface and selection of recombinant cells if the newly acquired DNA conveys antibiotic resistance to the transformed cells.

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Protocols for Transformation

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Videos

  1. TransformationVideo_thumb

    The Mechanism of Transformation with Competent Cells

    Transformation is the process by which bacteria are made to take up exogenous DNA. The word is derived from Griffith's discovery of a "transforming principle". Learn more about transformation and how it is used in cloning workflows.

  2. TransformationTips_VideoThumb_282x210_0616

    Tips for Successful Transformations with NEB Competent Cells

    Follow these tips to get superior results.

  3. TransformationProtocol_VideoThumb_282x210_0616

    How to Perform a Transformation with NEB Competent Cells

    Use this protocol for maximum transformation efficiency.

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