Features of Determining Thermodynamic Parameters of Formation of Nucleic Acid Complexes Using Thermal Denaturation with Fluorimetric Signal Detection


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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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