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DNA‐probes for the highly sensitive identification of single nucleotide polymorphism using single‐molecule spectroscopy
Author(s) -
Friedrich Achim,
Hoheisel Jörg D.,
Marmé Nicole,
Knemeyer Jens-Peter
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.03.031
Subject(s) - oligonucleotide , fluorescence , fluorescence spectroscopy , dna , nucleotide , single nucleotide polymorphism , chemistry , fluorescence correlation spectroscopy , single molecule experiment , guanosine , oligomer restriction , biophysics , hybridization probe , spectroscopy , molecule , biology , biochemistry , gene , physics , organic chemistry , quantum mechanics , genotype
This article presents a new, highly sensitive method for the identification of single nucleotide polymorphisms (SNPs) in homogeneous solutions using fluorescently labeled hairpin‐structured oligonucleotides (smart probes) and fluorescence single‐molecule spectroscopy. While the hairpin probe is closed, fluorescence intensity is quenched due to close contact between the chromophore and several guanosine residues. Upon hybridization to the respective target SNP sequence, contact is lost and the fluorescence intensity increases significantly. High specificity is achieved by blocking sequences containing mismatch with unlabeled oligonucleotides. Time‐resolved single‐molecule fluorescence spectroscopy enables the detection of individual smart probes passing a small detection volume. This method leads to a subnanomolar sensitivity for this single nucleotide specific DNA assay technique.