@article{oai:repo.qst.go.jp:00045401, author = {Michikawa, Yuichi and Suga, Tomo and Ishikawa, Atsuko and Ohtsuka, Yoshimi and Iwakawa, Mayumi and Imai, Takashi and 道川 祐市 and 菅 智 and 石川 敦子 and 荘司 好美 and 岩川 眞由美 and 今井 高志}, journal = {Oligonucleotide Array Sequence Analysis}, month = {}, note = {Haplotypes of multiple genes have been reported their association with risk of adverse skin reactions after radiotherapy in breast cancer patients (Int. J. Radiat. Oncol. Biol. 2007, 69, 685-693). In this study, a visible genotyping array device for a total of eight haplotypetagSNPs in two genes (PTTG1 and CD44) among above has been developed using previously established methodology (Anal. Sci. 2006, 22, 1537-1545). The device uses allele-specific extension reaction of covalently immobilized oligonucleotide primer to discriminate nucleotide at SNP site. Each primer oligonucleotide has 3'-end locked acid nucleotide modification to enhance its allelic specificity. Biotin-dUTPs provided in the reaction mixture are selectively incorporated only to the extenting oligonucleotides, enabling their use as tags for following streptavidin-conjugated alkaline phosphatase-mediated colored precipitation of substrates onto the surface of the array. Resulting colored spots on the device can be observed by naked eye and conveniently recorded by a digital camera that is commercially available at reasonable cost. Allelic discrimination of all immobilized oligonucleotides was validated using 45 reference individuals previously genotyped by MassARRAY system. Reliability of genotyping by this device was quantitatively assessed by calculating Silhouette score and resulting scores of all SNPs were proven to be beyond the cut off value of 0.65. The eight haplotype-tagSNPs can be simultaneously analyzed in a single reaction mixture within a couple of hours and multiple samples up to four can be analyzed simultaneously on a single device whose surface area is divided by a water-proof multiwell seal. This visible haplotype-tagSNP typing array device has intended benefits in convenient usage at reasonable cost and thus it would provide easy access for bedside clinical diagnosis of radiation sensitivity.}, pages = {3--14}, title = {Visible haplotype-tagSNP typing array device for human radiation sensitivity-associated genes}, year = {2008} }