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



: Ketomaki K, Hakala H, Kuronen O, Lonnberg H

Publisher: AMER CHEMICAL SOC

: 2003

Bioconjugate Chemistry

BIOCONJUGATE CHEMISTRY

: BIOCONJUGATE CHEM

: 14

: 4

: 811

: 816

: 6

: 1043-1802

DOI: https://doi.org/10.1021/bc0340058


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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