Label‐Free Voltammetric Detection of Products of Terminal Deoxynucleotidyl Transferase Tailing Reaction
2019
A label‐free approach that takes advantage of intrinsic
electrochemical activity of nucleobases has been applied to
study the products of terminal deoxynucleotidyl transferase
(TdT) tailing reaction. DNA homooligonucleotides A30, C30 and
T30 were used as primers for the tailing reaction to which a
dNTP – or a mixture of dNTPs – and TdT were added to form the
tails. Electrochemical detection enabled study of the tailing
reaction products created by various combinations of primers
and dNTPs, with pyrolytic graphite electrode (PGE) being
suitable for remarkably precise analysis of the length of
tailing reaction products. Furthermore, the hanging mercury
drop electrode (HMDE) was able to reveal formation of various
DNA structures, such as DNA hairpins and G‐quadruplexes, which
influence the behavior of DNA molecules at the negatively
charged surface of HMDE. Thus, the described approach proves to
be an excellent tool for studying the TdT tailing reactions and
for exploring how various DNA structures affect both the
tailing reactions and electrochemical behavior of DNA
oligonucleotides at electrode surfaces.
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