Electrochemical detection of nucleic base mismatches with ferrocenyl naphthalene diimide.
Electrochemical detection of nucleic base mismatches was attempted successfully with ferrocenyl naphthalene diimide (FND) in a model system with 20-meric double-stranded oligonucleotides with or without a mismatch(es). Thus, dA(20) or a 20-meric sequence of the lac Z gene was immobilized on a gold electrode and complementary oligonucleotides with different numbers of mismatches were allowed to hybridize in the presence of FND to give rise to an electrochemical signal. The signal intensity varied depending on the number of unpaired bases on the DNA duplex. From experiments with a quartz crystal microbalance, eight molecules of FND were found to bind to the 20-meric double-stranded oligos and this number decreased as the number of mismatches increased. These findings were further supported by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy. This novel method will be useful for the analysis of single-nucleotide polymorphisms present on human genes.
Yamashita
Kenichi
K
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka 812-8581, Japan.
Takagi
Makoto
M
Kondo
Hiroki
H
Takenaka
Shigeori
S
D020137:Base Pair Mismatch; D004563:Electrochemistry; D004926:Escherichia coli / Q000235:genetics; D005296:Ferrous Compounds; D007094:Imides; D009841:Oligonucleotides / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005296:Ferrous Compounds; D007094:Imides; D009841:Oligonucleotides; C406363:ferrocenylnaphthalene diimide