Gene Splicing by Overlap Extension: Tailor-Made Genes Using the Polymerase Chain Reaction

TLDR

Gene Splicing by Overlap Extension is a PCR-based technique that recombines DNA sequences without restriction sites and directly generates mutated fragments in vitro. By designing primers with overlapping sequences, PCR products share a common end that hybridizes, and DNA polymerase extends the overlap to produce a recombinant molecule. This approach is powerful, technically simple, and offers many advantages over conventional gene manipulation methods.

Abstract

Gene Splicing by Overlap Extension or "gene SOEing" is a PCR-based method of recombining DNA sequences without reliance on restriction sites and of directly generating mutated DNA fragments in vitro. By modifying the sequences incorporated into the 5'-ends of the primers, any pair of polymerase chain reaction products can be made to share a common sequence at one end. Under polymerase chain reaction conditions, the common sequence allows strands from two different fragments to hybridize to one another, forming an overlap. Extension of this overlap by DNA polymerase yields a recombinant molecule. This powerful and technically simple approach offers many advantages over conventional approaches for manipulating gene sequences.