A simple and efficient method for the oligonucleotide-directed mutagenesis using plasmid DNA template and phosphorothioate-modified nucleotide

Manabu Sugimoto, Nobuyoshi Esaki, Hidehiko Tanaka, Kenji Soda

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We have developed a simple and efficient method for oligonucleotide-directed mutagenesis with double-stranded (plasmid) DNA as a template. The template was simply and rapidly prepared by cell lysis and the following DNA denaturation with alkali. The chain elongation was performed with phosphorothioate-modified nucleotide at 37°C. After the selective digestion of original DNA with NciI and exonuclease III, the desired mutated gene was obtained at a high frequency (about 70%).

Original languageEnglish
Pages (from-to)309-311
Number of pages3
JournalAnalytical Biochemistry
Volume179
Issue number2
DOIs
Publication statusPublished - Jun 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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