Hybridization properties of support-bound oligonucleotides: the effect of the site of immobilization on the stability and selectivity of duplex formation.
Four 12-mer oligodeoxyribonucleotide sequences were immobilized to uniformly sized (50 microm) 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.
Ketomäki
Kaisa
K
Department of Chemistry, University of Turku, FIN-20014 Turku, Finland. kaisa.ketomaki@utu.fi
Hakala
Harri
H
Kuronen
Outi
O
Lönnberg
Harri
H
D020137:Base Pair Mismatch; D001665:Binding Sites; D008956:Models, Chemical; D015394:Molecular Structure; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry; D010316:Particle Size; D012680:Sensitivity and Specificity
D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes