2'-O-Methyl Oligoribonucleotide Analogs Used to Change the Temperature Characteristics of Immobilized Probes and to Enhance the Specificity of Hybridization


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The specificity of interactions between oligonucleotide probes immobilized in a biological microchip (biochip) and target DNA depends on a number of factors, with the main roles being played by the probe length and nucleotide sequence (GC composition primarily). The 2'-O-methyl oligoribonucleotides are known to form more stable duplexes with complementary DNA compared with 2'-deoxyribonucleotide probes. The study tested the possibility of using 2'-O-methyl oligoribonucleotides to increase the specificity of interactions between target DNA and probes immobilized in a biochip. Fluorophore-labeled target DNA was obtained via one-round asymmetric PCR with simultaneous incorporation of a fluorescent label in the PCR product. After hybridization with immobilized probes, fluorescent signals from gel pads with 2'-O-methyl oligoribonucleotide probes were, on average, two times higher than from those with 2'-deoxyribonucleotide probes, with the target DNA sequences and probe concentrations being the same. The increase in fluorescence intensity was greater in the case of perfect versus imperfect (mismatched) duplexes.

作者简介

A. Ikonnikova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: nased@biochip.ru
俄罗斯联邦, Moscow, 119991

S. Surzhikov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: nased@biochip.ru
俄罗斯联邦, Moscow, 119991

V. Pozhitnova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: nased@biochip.ru
俄罗斯联邦, Moscow, 119991

A. Zasedatelev

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: nased@biochip.ru
俄罗斯联邦, Moscow, 119991

T. Nasedkina

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

编辑信件的主要联系方式.
Email: nased@biochip.ru
俄罗斯联邦, Moscow, 119991

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