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

94 Scopus citations

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
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

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

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