TY - JOUR
T1 - Label-free voltammetric detection of products of terminal deoxynucleotidyl transferase tailing reaction
AU - Hermanová, Monika
AU - Havranová-Vidláková, Pavlína
AU - Ondráčková, Anna
AU - Kumar, Swathi Senthil
AU - Bowater, Richard
AU - Fojta, Miroslav
PY - 2019/2
Y1 - 2019/2
N2 -
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 A
30
, C
30
and T
30
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.
AB -
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 A
30
, C
30
and T
30
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.
KW - DNA electrochemistry
KW - label free
KW - nucleobase
KW - oligonucleotide tailing
KW - oxidation
KW - reduction
KW - terminal deoxynucleotidyl transferase
UR - http://www.scopus.com/inward/record.url?scp=85052462326&partnerID=8YFLogxK
U2 - 10.1002/elan.201800452
DO - 10.1002/elan.201800452
M3 - Article
AN - SCOPUS:85052462326
SN - 1040-0397
VL - 31
SP - 246
EP - 255
JO - Electroanalysis
JF - Electroanalysis
IS - 2
ER -