Twin probes as a novel tool for the detection of single-nucleotide polymorphisms.
Single-nucleotide polymorphisms (SNPs) are the most common form of DNA sequence variation. There is a strong interest from both academy and industry to develop rapid, sensitive and cost effective methods for SNP detection. Here we report a novel structural concept for DNA detection based on fluorescence dequenching upon hybridization. The so-called "twin probe" consists of a central fluorene derivative as fluorophore to which two identical oligonucleotides are covalently attached. This probe architecture is applied in homogeneous hybridization assays with subsequent fluorescence spectroscopic analysis. The bioorganic hybrid structure is well suited for sequence specific DNA detection and even SNPs are identified with high efficiency. Additionally, the photophysical properties of the twin probe were investigated. The covalent attachment of two single stranded oligonucleotides leads to strong quenching of the central fluorescence dye induced by the nucleobases. The twin probe is characterized by supramolecular aggregate formation accompanied by red-shifted emission and broad fluorescence spectra.
Ergen
Erhan
E
Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
Weber
Markus
M
Jacob
Josemon
J
Herrmann
Andreas
A
Müllen
Klaus
K
D001671:Biochemistry / Q000379:methods; D001710:Biotin / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry*; D005449:Fluorenes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D010777:Photochemistry / Q000379:methods; D020641:Polymorphism, Single Nucleotide
D015342:DNA Probes; D005449:Fluorenes; D005456:Fluorescent Dyes; D009841:Oligonucleotides; C041509:fluorene; D001710:Biotin