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PCR-mediated recombination and mutagenesis. SOEing together tailor-made genes
1Department of Biochemistry, College of Biological Sciences, St. Paul, MN 55108, USA.
Molecular Biotechnology
|April 1, 1995
Abstract:
Gene Splicing by Overlap Extension (gene SOEing) is a sequence-independent method for site-directed mutagenesis and/or recombination of DNA molecules. It is based on the idea that a PCR product can be engineered by adding or changing sequences at its ends so that the product can itself be used to prime DNA synthesis in a subsequent overlap-extension reaction to create mutant or recombinant molecules. As the engineered genes are created in vitro without reliance on host organisms or restriction sites, gene SOEing provides a powerful and versatile tool for genetic investigation and engineering.