Open AccessGene Splicing by Overlap Extension: Tailor-Made Genes Using the Polymerase Chain Reaction
Author(s) -
Robert M. Horton,
Zeiling Cai,
Steffan M Ho,
Larry R. Pease
Publication year - 2013
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114017
Subject(s) - overlap extension polymerase chain reaction , polymerase chain reaction , genetics , biology , gene , polymerase , rna splicing , dna , inverse polymerase chain reaction , computational biology , sequence (biology) , restriction enzyme , microbiology and biotechnology , nested polymerase chain reaction , rna
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.