Team:Utah State/Results

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Electroporation Transformation of E. Coli

Transformations are any procedure used to insert DNA into a bacteria (if you use a virus, the term becomes transfection). Electroporation uses a pulse of electricity to disrupt the cell membrane and create holes that would allow the DNA to enter the cell. Cells need to be made competent before doing this procedure, in order for them to efficiently take up the DNA. Transformations generally utilize millions to billions of cells and DNA molecules and, for a transformation to be successful, only one molecule of DNA needs to enter into one cell, which then grows into a colony. One issue with transformations is selecting and verifying which colonies have the desired DNA. This is usually done using a marker, a characteristic possessed by cells that have the DNA (or lost by those cells) that distinguishes it from the rest of the colonies that grow up. Commonly, this is the expression of an antibiotic resistance gene included on the transformed DNA, which allows only the cells that have taken up the DNA to survive on a plate in the presence of that antibiotic. Sometimes pigment producing or fluorescent/luminescent proteins can also be used in place of antibiotic resistance to allow visual determination of transformed colonies. Other ways of selection exist, but will not be discussed here.