title: Tag/hybridization-based sensitive detection of polymerase chain reaction products.
abstract: The polymerase chain reaction (PCR) is an important technology to amplify a single copy or a few copies of DNA segment in genomic DNAs, visualizing the segment as DNA fragment. Thus, PCR is frequently used in various examinations such as detection of bacteria and fungi in the food industry. Here, we report a simple and sensitive method for detection of PCR products using single-strand tag sequence and hybridization of the tag sequence to the complementary tag sequence immobilized on solid material (STH). The detection sensitivity was found to be at least 50 times higher than electrophoresis/ethidium bromide (EtBr) visualization for approximately a 500-bp fragment and higher than the ordinary hybridization, that is, hybridization of denatured PCR product to probe sequence immobilized on solid material.
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan. Electronic address: kniwa@ngk.co.jp.
authors: Kousuke
authors: K
authors: Niwa
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan.
authors: Akinobu
authors: A
authors: Oribe
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan.
authors: Hidemasa
authors: H
authors: Okumura
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan.
authors: Masahiro
authors: M
authors: Shimono
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan.
authors: Kenkichi
authors: K
authors: Nagai
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan.
authors: Toshikazu
authors: T
authors: Hirota
authors: Tsukuba Gene Technology Laboratories, Tsuchiura, Ibaraki 300-0873, Japan.
authors: Hiroshi
authors: H
authors: Yasue
authors: Future Technology Management Center, Corporate R&D, NGK Insulators, Mizuho, Nagoya 467-8530, Japan; Graduate School of Biomedical Engineering, Tohoku University, Aoba, Sendai 980-8579, Japan.
authors: Mitsuo
authors: M
authors: Kawase
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics; D006801:Humans; D057230:Limit of Detection; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA
keywords: DNA microarray; Fluorescence label; Hybridization; Non-nucleic acid spacer; Single-stranded DNA tag