Further development of multiplex single nucleotide polymorphism typing method, the DigiTag2 assay
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作者:
Nishida, Nao
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Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, JapanUniv Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
Nishida, Nao
[1
]
Tanabe, Tetsuya
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机构:Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
Tanabe, Tetsuya
Takasu, Miwa
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机构:Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
Takasu, Miwa
Suyama, Akira
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机构:Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
Suyama, Akira
Tokunaga, Katsushi
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机构:Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
Tokunaga, Katsushi
机构:
[1] Univ Tokyo, Grad Sch Med, Dept Human Genet, Bunkyo Ku, Tokyo 1130033, Japan
[2] Olympus Corp, Bio Business Div, Hachioji, Tokyo 1928512, Japan
[3] Univ Tokyo, Grad Sch Arts & Sci, Dept Life Sci, Meguro Ku, Tokyo 1538902, Japan
A number of single nucleotide polymorphisms (SNPs) are considered to be candidate susceptibility or resistance genetic factors for multifactorial disease. Genome-wide searches for disease susceptibility regions followed by high-resolution mapping of primary genes require cost-effective and highly reliable technology. To accomplish successful and low-cost typing for candidate SNPs, new technologies must be developed. We previously reported a multiplex SNP typing method, designated the DigiTag assay, that has the potential to analyze nearly any SNP with high accuracy and reproducibility. However, the DigiTag assay requires multiple washing steps in manipulation and uses genotyping probes modified with biotin for each target SNP. Here we describe the next version of the assay, DigiTag2, which works with simple protocols and uses unmodified genotyping probes. We investigated the feasibility of the DigiTag2 assay by genotyping 96 target SNPs spanning a 610-kb region of human chromosome 5. The DigiTag2 assay is suitable for genotyping an intermediate number of SNPs (tens to hundreds of sites) with a high conversion rate (> 90%), high accuracy, and low cost. (c) 2007 Elsevier Inc. All rights reserved.