Simple detection of point mutations in DNA oligonucleotides using SYBR Green I

Tatsuo Maruyama, Tomoaki Takata, Hirofumi Ichinose, Lian Chun Park, Noriho Kamaiya, Masahiro Goto

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

A novel and simple method for detection of mutations in DNA oligonucleotides using a double-stranded DNA specific dye (SYBR Green I) is reported. The SYBR Green I is bound specifically with a duplex DNA oligonucleotide (intercalation). This intercalation induces fluorescent emission at 525 nm with excitation at 494 nm. The fluorescence intensity of mismatched oligonucleotides (40-mer) decreases (by more than 13%) in comparison with the perfectly matched oligonucleotides. Moreover, fluorescence measurement of the SYBR Green I can distinguish various types of single-base mismatches, except for the T-G terminal mismatch. The addition of 20% (v/v) formamide, however, to an oligonucleotide solution improved the sensitivity of detection and also enabled the detection of the T-G terminal-mismatch. This detection method requires only a normal fluorescence spectrophotometer, an inexpensive dye and just 50 pmol of sample DNA.

Original languageEnglish
Pages (from-to)1637-1641
Number of pages5
JournalBiotechnology letters
Volume25
Issue number19
DOIs
Publication statusPublished - Oct 1 2003

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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