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Reliable and Fast Allele-Specific Extension of 3'-LNA Modified Oligonucleotides Covalently Immobilized on a Plastic Base, Combined with Biotin-dUTP Mediated Optical Detection
https://repo.qst.go.jp/records/44742
https://repo.qst.go.jp/records/447426ce8fe54-8734-482e-a146-4ab7cf94281e
Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2006-12-15 | |||||
タイトル | ||||||
タイトル | Reliable and Fast Allele-Specific Extension of 3'-LNA Modified Oligonucleotides Covalently Immobilized on a Plastic Base, Combined with Biotin-dUTP Mediated Optical Detection | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
アクセス権 | ||||||
アクセス権 | metadata only access | |||||
アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
著者 |
Michikawa, Yuichi
× Michikawa, Yuichi× Fujimoto, Kentaro× Kinoshita, Kenji× Kawai, Seiko× Sugahara, Keisuke× Suga, Tomo× Ootsuka, Yoshimi× Fujiwara, Kazuhiko× Iwakawa, Mayumi× Imai, Takashi× 道川 祐市× 川井 聖子× 菅原 圭亮× 菅 智× 荘司 好美× 岩川 眞由美× 今井 高志 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | In the present work, a convenient microarray SNP typing system has been developed using a plastic base that covalently immobilizes amino-modified oligonucleotides. Reliable SNP allele discrimination was achieved by using allelic specificity-enhanced enzymatic extension of immobilized oligonucleotide primer, with a locked nucleic acid (LNA) modification at the SNP-discriminating 3'-end nucleotide. Incorporation of multiple biotin-dUTP molecules during primer extension, followed by binding of alkaline phosphatase-conjugated streptavidin, allowed optical detection of the genotyping results through precipitation of colored alkaline phosphatase substrates onto the surface of the plastic base. Notably, rapid primer extension was demonstrated without a preliminary annealing step of double-stranded template DNA, allowing overall processes to be performed within a couple of hours. Simultaneous evaluation of three SNPs in the genes TGFB1, SOD2 and APEX1, previously investigated for association with radiation sensitivity, in 25 individuals has shown perfect assignment with data obtained by another established technique (Massarray system). | |||||
書誌情報 |
Analytical Sciences 巻 22, 号 12, p. 1537-1545, 発行日 2006-12 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 0910-6340 |