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

G. Y. Shevelev, M. R. Kabilov, A. A. Lomzov, I. S. Dovydenko, D. V. Pyshnyi

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)684-698
Number of pages15
JournalRussian Journal of Bioorganic Chemistry
Volume45
Issue number6
DOIs
Publication statusPublished - 1 Nov 2019

Keywords

  • fluorimetry
  • melting temperature of DNA complexes
  • nucleic acid complexes
  • oligonucleotides
  • thermodynamic analysis

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