Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching

Tatsuo Maruyama, Toshimitsu Shinohara, Takuya Hosogi, Hirofumi Ichinose, Noriho Kamiya, Masahiro Goto

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Guanine quenching of a fluorescence-labeled DNA probe is a powerful tool for detecting a mutation in a targeted site of a DNA strand. However, a different guanine adjacent to a targeted site can interfere with detection of a point mutation, resulting in unsatisfactory sensitivity. In the current study, we developed a simple method to improve sensitivity of the guanine quenching method using a masking DNA oligonucleotide. The simple addition of a masking DNA oligonucleotide was found to mask the interference of a different guanine in a target oligonucleotide on fluorescence and to enhance difference in the quenching ratio between wild-type and mutant oligonucleotides. Based on this strategy, we succeeded in discriminating various mutations from the wild-type YMDD motif of the hepatitis B virus DNA polymerase gene using guanine quenching with a masking oligonucleotide.

Original languageEnglish
Pages (from-to)8-14
Number of pages7
JournalAnalytical Biochemistry
Volume354
Issue number1
DOIs
Publication statusPublished - Jul 1 2006

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All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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