Visible Genotype Sensor Array 英文参考文献.docVIP

Sensors 2008, 8, 2722-2735 sensors ISSN 1424-8220 ? 2008 by MDPI /sensors Review Visible Genotype Sensor Array Yuichi Michikawa*, Tomo Suga, Yoshimi Ohtsuka, Izumi Matsumoto, Atsuko Ishikawa, Kenichi Ishikawa, Mayumi Iwakawa, and Takashi Imai RadGenomics Project, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba, 263-8555, Japan; E-mails: tomo@nirs.go.jp, y_otsuka@nirs.go.jp, spring@nirs.go.jp, a_ishi@nirs.go.jp, k_ishi@nirs.go.jp, mayumii@nirs.go.jp, imait@nirs.go.jp * Author to whom correspondence should be addressed; E-mail: y_michi@nirs.go.jp; Telephone: +011-81-43-206-3072 Fax: +011-81-43-206-6267; Received: 25 January 2008 / Accepted: 15 April 2008 / Published: 17 April 2008 Abstract: A visible sensor array system for simultaneous multiple SNP genotyping has been developed using a new plastic base with specific surface chemistry. Discrimination of SNP alleles is carried out by an allele-specific extension reaction using immobilized oligonucleotide primers. The 3’-ends of oligonucleotide primers are modified with a locked nucleic acid to enhance their efficiency in allelic discrimination. Biotin-dUTPs included in the reaction mixture are selectively incorporated into extending primer sequences and are utilized as tags for alkaline phosphatase-mediated precipitation of colored chemical substrates onto the surface of the plastic base. The visible precipitates allow immediate inspection of typing results by the naked eye and easy recording by a digital camera equipped on a commercial mobile phone. Up to four individuals can be analyzed on a single sensor array and multiple sensor arrays can be handled in a single operation. All of the reactions can be performed within one hour using conventional laboratory instruments. This visible genotype sensor array is suitable for “focused genomics” that follows “comprehensive g