Ferrocenylnaphthalene diimide-based electrochemical detection of methylated gene.
Ferrocenylnaphthalene diimide (FND)-based electrochemical hybridization assay was applied to the detection of methylated cytosine of DNA using the products obtained after treatment with bisulfite followed by polymerase chain reaction (PCR), where unmethylated cytosine is converted to thymine and methylated one to cytosine. Twenty-meric DNA probes for the methylated (cytosine) and unmethylated (thymine) types of the part of the promoter region of cyclin D-dependent protein kinase inhibitor, p16, gene (p16(Ink4a)) were used to be immobilized on the electrochemical array (ECA) chip. Using 1 microL of 10 ng/microL of methylated sample obtained from the methylation-specific PCR of methylated genome containing 10-times excess of unmethylated one, the methylated PCR sample could be detected by the identical electrochemical signals from the two DNA probes under the settled optimum hybridization conditions.
Sato
Shinobu
S
Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Iizuka 820-8502, Japan.
Hokazono
Koji
K
Irie
Tatsuya
T
Ueki
Takashi
T
Waki
Michinori
M
Nojima
Takahiko
T
Kondo
Hiroki
H
Takenaka
Shigeori
S