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Intercalating Nucleic Acids with Pyrene Nucleotide Analogues as Next‐Nearest Neighbors for Excimer Fluorescence Detection of Single‐Point Mutations under Nonstringent Hybridization Conditions
Author(s) -
Christensen Ulf B.,
Pedersen Erik B.
Publication year - 2003
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/hlca.200390165
Subject(s) - chemistry , pyrene , excimer , nucleic acid , fluorescence , intercalation (chemistry) , quenching (fluorescence) , point mutation , nucleic acid thermodynamics , nucleobase , nucleotide , stereochemistry , photochemistry , mutant , dna , biochemistry , organic chemistry , base sequence , gene , physics , quantum mechanics
Fluorescence and hybridization specificity is reported for intercalating nucleic acids (INAs), which are here oligodeoxynucleotides (ODNs) synthesized with insertions using ( S )‐1‐[bis(4‐methoxyphenyl)(phenyl)methoxy]‐3‐[(pyren‐1‐yl)methoxy]propan‐2‐ol phosphoramidites. It is shown that an INA with two insertions placed as next‐nearest neighbors can be used for discrimination between nucleic acids and their single‐point mutants. Quenching of an excimer band at 480 nm is observed upon hybridizing to a complementary sequence, whereas the excimer band is present when the nucleobase pair between the two pyrene moieties is mismatched. It is the first example of a solution based on fluorescence detection of single‐point mutants that uses excimer formation and does not rely on stringent hybridization conditions. Furthermore, it is shown that INAs with pyrene insertions retain their sequence specificity in thermal melting.