Features of Determining Thermodynamic Parameters of Formation of Nucleic Acid Complexes Using Thermal Denaturation with Fluorimetric Signal Detection
- Authors: Shevelev G.Y.1,2, Kabilov M.R.1, Lomzov A.A.1,2, Dovydenko I.S.1, Pyshnyi D.V.1,2
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Affiliations:
- Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
- Novosibirsk State University
- Issue: Vol 45, No 6 (2019)
- Pages: 684-698
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/229301
- DOI: https://doi.org/10.1134/S1068162019060359
- ID: 229301
Cite item
Abstract
In this work, the possibility of precise physicochemical analysis of formation of nucleic acid complexes has been demonstrated using thermal cyclers with fluorescence signal detection. Individual features of heating blocks of various thermal cyclers have been identified. A nonlinear correlation between fluorescence signal intensity and concentration of fluorescent dyes (5,6)-carboxyfluorescein and Rhodamine B has been identified. Using UV and fluorescence melting, the comparison of melting curves at various concentration of full-complementary and mismatched (A/A and C/A) complexes has been conducted. The experimental values of thermodynamic parameters of complex formation for full-complementary and mismatched complexes have been obtained.
About the authors
G. Y. Shevelev
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences; Novosibirsk State University
Author for correspondence.
Email: metatezis@gmail.com
Russian Federation, Novosibirsk, 630090; Novosibirsk, 630090
M. R. Kabilov
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
Email: metatezis@gmail.com
Russian Federation, Novosibirsk, 630090
A. A. Lomzov
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences; Novosibirsk State University
Email: metatezis@gmail.com
Russian Federation, Novosibirsk, 630090; Novosibirsk, 630090
I. S. Dovydenko
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences
Email: metatezis@gmail.com
Russian Federation, Novosibirsk, 630090
D. V. Pyshnyi
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences; Novosibirsk State University
Email: metatezis@gmail.com
Russian Federation, Novosibirsk, 630090; Novosibirsk, 630090
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