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Site-directed mutagenesis is the process of intentionally introducing a specific mutation into a specific gene or DNA sequence. Usually site-directed mutagenesis is a polymerase chain reaction (PCR)-based approach. Often a primer that contains a mutation is used or regions of a gene are added or deleted through PCR. Once the mutated gene is created, it is inserted into a vector, which is a DNA molecule for transport of DNA into a host cell, to create a circularized piece of DNA called a plasmid. The plasmid is added to host cells, usually bacteria, where it is replicated and then purified out of the cells. The mutant gene can then be added to cells to observe the resultant phenotype or can be used to generate recombinant protein containing the mutation. Site-directed mutagenesis has multiple uses from studying gene function to drug development to creating biofuel-producing bacteria. (Credit: Brooke Anderson-White)
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