Splicing by Overlap Extension PCR to Obtain Hybrid DNA Products

Methods Mol Biol. 2016:1373:43-9. doi: 10.1007/7651_2014_182.

Abstract

Genetic manipulation of bacteria often requires the joining together of more than one DNA segment to form a hybrid DNA molecule. This can be accomplished by PCR followed by restriction endonuclease digestions and ligations. However, this approach can often become laborious and expensive. Here is described a well-established method for using primer design and PCR to obtain hybrid products for use in cloning vectors, mutagenesis protocols, and other applications.

Keywords: Gene splicing; Mutagenesis; PCR fusion; Polymerase chain reaction.

MeSH terms

  • DNA Primers / genetics
  • Genetic Vectors / genetics
  • Humans
  • Mutagenesis, Site-Directed / methods
  • Nucleic Acid Hybridization / methods*
  • Polymerase Chain Reaction / methods*
  • RNA Splicing / genetics*
  • Staphylococcus / genetics*
  • Staphylococcus / pathogenicity

Substances

  • DNA Primers