DNA sensor: A novel electrochemical gene detection method using carbon electrode immobilized DNA probes |
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Authors: | Koji Hashimoto Keiko Miwa Masanori Goto Yoshio Ishimori |
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Institution: | Materials and Devices Research Laboratories, Research and Development Center, Toshiba Corporation , 1, Komukai Toshiba-cho, Saiwai-ku, Kawasaki, 210, Japan |
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Abstract: | Abstract The authors have developed a novel, rapid, convenient, and specific gene detection method, named the ‘DNA sensor,’ using a graphite electrode loaded with DNA probes. Synthesized oligonucleotide (5-TGCAGTTCCGGTGGCTGATC-3′) complementary to oncogene v-myc was employed for a model probe. The oligonucleotide was chemically adsorbed on a basal plane pyrolytic graphite (BPPG) electrode. The sensor was able to be applied to a hybridization reaction (40°C) in a linearized pVM623 solution carrying the Pst I fragment of v-myc (1.5 kbp). After the hybridization reaction, the sensor was immersed into an acridine orange solution (1 μM) and washed with a phosphate buffer (pH 7.0). Acridine orange intercalated between base pairs of the formed double stranded DNAs on the electrode. The anodic peak potential of acridine orange that interacted with the DNAs on the electrode was measured. The positive shift of the peak potential increased in proportional to the pVM623 concentration in the hybridization reaction. 10?1 g/ml of pVM623 was able to be detected in the buffer solution using the sensor. This gene detection was completed within an hour. |
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