In Vitro Recombination and Mutagenesis of DNA: SOEing Together Tailor-Made Genes

Abstract

Gene Splicing by Overlap Extension (gene SOEing) provides a powerful method of recombining sequences without depending on restriction sites or ligase, and a simple, generally applicable way of using polymerase chain reaction (PCR) to perform site-directed mutagenesis in vitro. This technique allows even those with minimal molecular biology expertise to generate quickly genetic constructs that might otherwise have been impractical using only the older (restriction enzymebased) technology.