Single-mismatch detection using gold-quenched fluorescent oligonucleotides

Nat Biotechnol. 2001 Apr;19(4):365-70. doi: 10.1038/86762.

Abstract

Here we describe a hybrid material composed of a single-stranded DNA (ssDNA) molecule, a 1.4 nm diameter gold nanoparticle, and a fluorophore that is highly quenched by the nanoparticle through a distance-dependent process. The fluorescence of this hybrid molecule increases by a factor of as much as several thousand as it binds to a complementary ssDNA. We show that this composite molecule is a different type of molecular beacon with a sensitivity enhanced up to 100-fold. In competitive hybridization assays, the ability to detect single mismatch is eightfold greater with this probe than with other molecular beacons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Pair Mismatch*
  • Base Sequence
  • Binding, Competitive
  • DNA, Single-Stranded / chemistry
  • Dose-Response Relationship, Drug
  • Fluorescent Dyes / chemistry*
  • Genetic Techniques*
  • Gold / chemistry
  • Molecular Sequence Data
  • Nucleic Acid Hybridization*
  • Oligonucleotides / chemistry*
  • Protein Binding
  • Protein Conformation
  • Time Factors

Substances

  • DNA, Single-Stranded
  • Fluorescent Dyes
  • Oligonucleotides
  • Gold