Gene splicing by overlap extension: Tailor-made genes using the polymerase chain reaction

R. M. Horton, Z. Cai, S. N. Ho, Larry R Pease

Research output: Contribution to journalArticle

1090 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)528-535
Number of pages8
JournalBioTechniques
Volume8
Issue number5
StatePublished - 1990

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Clinical Biochemistry

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