Electrochemical analysis of template-free DNA synthesis: Lowering pH and the active component of the impedance spectrum

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• 作者：M. A. Gutorov ; V. V. Emets ; A. L. Klyuev…
• 刊名：Protection of Metals and Physical Chemistry of Surfaces
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：52
• 期：4
• 页码：684-688
• 全文大小：499 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Tribology, Corrosion and Coatings
  Materials Science
  Metallic Materials
  Inorganic Chemistry
  Industrial Chemistry and Chemical Engineering
  Characterization and Evaluation Materials
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：2070-206X
• 卷排序：52

文摘

The process of template-free DNA synthesis was detected in two ways: by measuring the pH of the solution by a semiconductor sensor and by measuring the conductivity in the recording of impedance spectra. Synthesis was carried out without using template DNA, with only two enzymes being involved in the reaction: DNA polymerase and nicking endonuclease (nickase) in the presence of deoxynucleotide triphosphates. Previously, Purushothaman et al., by applying the recording of the results of template-directed synthesis with a pH-sensitive field-effect transistor, showed that protons are released into solution in the incorporation of nucleotides [4]. Regarding this, it was important to establish that the same release of protons into solution occurred in the template-free synthesis as in the template-directed synthesis and to identify the changes by measuring the conductivity of the solution using impedance spectra. It was found that the template-free synthesis was accompanied by the generation of protons (ΔpH is ~1.5 pH at an initial concentration of deoxynucleotide triphosphates of 150 µM) and a decrease in the value of active impedance component by ~25% of the initial value. The effect of a decrease in the active impedance component was explained as being due to an increasing conductivity of medium due to a growth in the concentration of protons.Original Russian Text © M.A. Gutorov, V.V. Emets, A.L. Klyuev, V.N. Andreev, A.N. Reshetilov, 2016, published in Fizikokhimiya Poverkhnosti i Zashchita Materialov, 2016, Vol. 52, No. 4, pp. 431–435.