Preliminary evaluation of electrochemical PNA array for detection of single base mismatch mutations.
We report here a preliminary evaluation of a microfabricated disposable-type peptide nucleic acid (PNA) array, with a 20-channel electrode, for the detection of cancer gene c-Ki-ras. Synthetic 15-mer PNA probes complimentary to ras sequence modified with cysteine were immobilized on the gold electrodes on the array. The electrochemical PNA array was reacted with 20-mer oligonucleotide target or 128 bp PCR product for 1 h. The anodic current derived from an electrochemically active DNA binder Hoechst 33258 was measured using the PNA array in the 50 microL of reaction chamber. The anodic current from Hoechst 33258 increased with increasing the concentration of PCR product of ras gene in the range from 10(11) to 10(15) copy mL(-1). The single base mismatch mutations of c-Ki-ras/61 were also detected using the electrochemical PNA array.
Hashimoto
K
K
Corporate Research and Development Center, Toshiba, 1, Komukai, Toshiba-cho, Saiwai-ku, Kawasaki, 212-8582, Japan. kouji.hashimoto@toshiba.co.jp
Ishimori
Y
Y
D020137:Base Pair Mismatch; D001483:Base Sequence; D015342:DNA Probes; D004563:Electrochemistry / Q000379:methods*; D011905:Genes, ras; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D020135:Peptide Nucleic Acids / Q000737:chemistry*
D015342:DNA Probes; D020135:Peptide Nucleic Acids