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Hybridization properties of support-bound oligonucleotides: The effect of the site of immobilization on the stability and selectivity of duplex formation




TekijätKetomaki K, Hakala H, Kuronen O, Lonnberg H

KustantajaAMER CHEMICAL SOC

Julkaisuvuosi2003

Lehti: Bioconjugate Chemistry

Tietokannassa oleva lehden nimiBIOCONJUGATE CHEMISTRY

Lehden akronyymiBIOCONJUGATE CHEM

Vuosikerta14

Numero4

Aloitussivu811

Lopetussivu816

Sivujen määrä6

ISSN1043-1802

DOIhttps://doi.org/10.1021/bc0340058


Tiivistelmä
Four 12-mer oligodeoxyribonucleotide sequences were immobilized to uniformly sized (50 mum) polymer particles through C5-tethered thymine and N-4-tethered cytosine bases at four different sites in each sequence. The effect of the site of immobilization on the efficiency and selectivity of hybridization of the particle-bound probes was quantified by a sandwich-type assay based on a time-resolved fluorometric measurement of an oligonucleotide probe labeled with a photoluminescent europium(III) chelate directly from the surface of a single particle. Immobilization through a base in the central part of the sequence was observed to destablize the duplex more markedly than tethering through a terminal base. The effect of a one-base mismatch on the duplex stability increased with the increasing distance from the site of immobilization.


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