2'-O-Methyl Oligoribonucleotide Analogs Used to Change the Temperature Characteristics of Immobilized Probes and to Enhance the Specificity of Hybridization
- Авторы: Ikonnikova A.Y.1, Surzhikov S.A.1, Pozhitnova V.O.1, Zasedatelev A.S.1, Nasedkina T.V.1
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Учреждения:
- Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
- Выпуск: Том 63, № 6 (2018)
- Страницы: 876-879
- Раздел: Molecular Biophysics
- URL: https://journals.rcsi.science/0006-3509/article/view/152780
- DOI: https://doi.org/10.1134/S0006350918060118
- ID: 152780
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Аннотация
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.
Об авторах
A. Ikonnikova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: nased@biochip.ru
Россия, Moscow, 119991
S. Surzhikov
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: nased@biochip.ru
Россия, Moscow, 119991
V. Pozhitnova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: nased@biochip.ru
Россия, Moscow, 119991
A. Zasedatelev
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: nased@biochip.ru
Россия, Moscow, 119991
T. Nasedkina
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Автор, ответственный за переписку.
Email: nased@biochip.ru
Россия, Moscow, 119991
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