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NUCLEIC ACID HYBRIDIZATION ACCOMPANIED WITH EXCIMER FORMATION FROM TWO PYRENE‐LABELED PROBES
Author(s) -
Ebata Katsuyoshi,
Masuko Masayuki,
Ohtani Hiroyuki,
KashiwasakeJibu Masaki
Publication year - 1995
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1995.tb09144.x
Subject(s) - pyrene , excimer , oligonucleotide , molecular beacon , fluorescence , chemistry , nucleic acid , monomer , molecular probe , nucleic acid thermodynamics , biophysics , photochemistry , biochemistry , dna , optics , base sequence , organic chemistry , biology , polymer , physics
— We developed a novel nucleic acid hybridization method based on excimer formation. We used two different 16‐mer oligonucleotide probes that had a combined continuous‐sequence run that was complementary to a target 32‐mer. Prior to hybridization, the adjacent terminal ends (i.e. the 3'‐terminal of one probe and the 5'‐terminal of the other probe) were each labeled with one pyrene residue. When these probes simultaneously hybridized to the target, a 495 nm broad fluorescence band was produced. The intensity of this band increased as the intensity of the pyrene monomer bands decreased, indicating that the 495 nm band was attributed to the pyrene excimer. The excimer fluorescence, easily differentiated from the monomer bands for emission wavelength, opens up a new way to perform homogeneous hybridization assays and in vivo imaging of nucleic acids.