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

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.

Keywords

Overlap extension polymerase chain reactionPolymerase chain reactionGeneticsBiologyPolymeraseGeneRNA splicingInverse polymerase chain reactionDNASequence (biology)Computational biologyRecombinant DNAMolecular biologyNested polymerase chain reactionRNA

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Publication Info

Year: 2013
Type: article
Volume: 54
Issue: 3
Pages: 129-133
Citations: 1303
Access: Closed

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APA Style

                            
                                
                                    Robert M. Horton, 
                                
                                    Zeling Cai, 
                                
                                    Steffan N. Ho
                                
                                et al.
                            
                            (2013). 
                            Gene Splicing by Overlap Extension: Tailor-Made Genes Using the Polymerase Chain Reaction. 
                            BioTechniques
                            , 54
                            (3)
                            , 129-133.
                            https://doi.org/10.2144/000114017
                        

Identifiers

DOI: 10.2144/000114017