A simple electroanalysis of polyG RNA in mixtures with 3′,5′-cyclic guanosine monophosphate achieved by selective desorption of the monomers from the electrode surface

• Published in: Journal of electroanalytical chemistry (Lausanne, Switzerland) Vol. 901; p. 115773
• Main Authors: Hesko, Ondrej, Fojta, Miroslav, Špaček, Jan
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.11.2021; Elsevier Science Ltd
• Subjects: 3′,5′-cGMP; Deionization; Desorption; Electrochemistry; Electrodes; Electrolytic analysis; Guanosines; High temperature; Monomers; Nucleotides; Oligonucleotides; Oxidation; Polymerization; Pyrolytic graphite; Pyrolytic graphite electrode; RNA; Electrochemistry; 3′,5′-cGMP; Desorption; RNA; Pyrolytic graphite electrode; Oligonucleotides
• ISSN: 1572-6657; 1873-2569
• DOI: 10.1016/j.jelechem.2021.115773

•A simple label-free technique for analysis of nucleic acid polymerization mixtures.•The technique is based on selective removal of interfering monomers from electrode.•Monomers are removed by treatment with a surfactant or water at elevated temperature. Previously it has been shown that cyclic nucleoside monophosphates can spontaneously polymerize to form RNA oligonucleotides under conditions simulating prebiotic conditions on Archean Earth. The most efficient polymerization was documented with 3′,5′-cyclic guanosine monophosphate (cGMP). In this work a method for fast detection of short polyG RNAs present in a large overabundance of cGMP, modeling conditions in the non-enzymatic nucleotide polymerization mixtures, is presented. The method is based on electrochemical measurements of guanine (G) oxidation signals yielded by RNA oligomers adsorbed onto the surface of a pyrolytic graphite electrode (PGE). To avoid false positive results arising from the G oxidation signals due to co-adsorbed cGMP, a method for selective removal of the monomers from the electrode surface has been devised. In the first step, both cGMP and RNAs are co-adsorbed onto the PGE surface. In the second step, the cGMP is selectively desorbed using treatments in solutions of different tested surfactants (SDS, Tween 20 or Triton X-100), or by washing in deionized water at elevated temperature. We show that this new approach is suitable for selective analysis of products of polymerization reactions from mixtures of their building blocks.