Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis

Gene. 1989 Mar 15;76(1):161-6. doi: 10.1016/0378-1119(89)90018-8.


The polymerase chain reaction catalyzed by Taq DNA polymerase has been used for site-specific mutagenesis. The amplification was primed by two oligodeoxyribonucleotides complementary to insulin receptor cDNA. To direct the synthesis of mutant DNA, mismatches were introduced into one of the primers. Six different mutations were constructed by this technique. Of twelve clones whose sequences were determined, ten (83%) had the correct sequence. This technique, which does not require the use of single-stranded DNA templates, provides a simple and efficient approach to site-specific mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Catalysis
  • Chemical Phenomena
  • Chemistry
  • Cloning, Molecular
  • DNA, Recombinant
  • DNA-Directed DNA Polymerase / metabolism*
  • Gene Amplification
  • Genetic Vectors
  • Mutation*
  • Oligodeoxyribonucleotides / biosynthesis
  • Oligodeoxyribonucleotides / genetics
  • Plasmids
  • Receptor, Insulin / genetics
  • Taq Polymerase
  • Templates, Genetic


  • DNA, Recombinant
  • Oligodeoxyribonucleotides
  • Receptor, Insulin
  • Taq Polymerase
  • DNA-Directed DNA Polymerase