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

Hiroko Kadowaki, Takashi Kadowaki, Fredric E. Wondisford, Simeon I. Taylor

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)161-166
Number of pages6
JournalGene
Volume76
Issue number1
DOIs
StatePublished - Mar 15 1989

Fingerprint

Taq Polymerase
Site-Directed Mutagenesis
Polymerase Chain Reaction
Oligodeoxyribonucleotides
Single-Stranded DNA
Insulin Receptor
Complementary DNA
Clone Cells
Mutation
DNA

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

  • Recombinant DNA
  • cloning
  • gene amplification
  • insulin receptor
  • oligodeoxyribonucleotide primers
  • plasmid

Cite this

Kadowaki, Hiroko ; Kadowaki, Takashi ; Wondisford, Fredric E. ; Taylor, Simeon I. / Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis. In: Gene. 1989 ; Vol. 76, No. 1. pp. 161-166.
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Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis. / Kadowaki, Hiroko; Kadowaki, Takashi; Wondisford, Fredric E.; Taylor, Simeon I.

In: Gene, Vol. 76, No. 1, 15.03.1989, p. 161-166.

Research output: Contribution to journalArticle

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