A1 Refereed original research article in a scientific journal
Hybridization properties of support-bound oligonucleotides: The effect of the site of immobilization on the stability and selectivity of duplex formation
Authors: Ketomaki K, Hakala H, Kuronen O, Lonnberg H
Publisher: AMER CHEMICAL SOC
Publication year: 2003
Journal:Bioconjugate Chemistry
Journal name in sourceBIOCONJUGATE CHEMISTRY
Journal acronym: BIOCONJUGATE CHEM
Volume: 14
Issue: 4
First page : 811
Last page: 816
Number of pages: 6
ISSN: 1043-1802
DOI: https://doi.org/10.1021/bc0340058
Abstract
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.
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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