Genotyping by allele-specific L-DNA-tagged PCR

被引：32

作者：

Hayashi, Gosuke ^{[1
]}

Hagihara, Masaki ^{[1
]}

Nakatani, Kazuhiko ^{[1
]}

机构：

[1] Osaka Univ, Inst Sci & Ind Res, Dept Regulatory Bioorgan Chem, Ibaraki 5670047, Japan

来源：

JOURNAL OF BIOTECHNOLOGY | 2008年 / 135卷 / 02期

关键词：

single nucleotide polymorphism (SNP) typing; allele-specific PCR; L-DNA; surface plasmon resonance (SPR) imaging;

D O I：

10.1016/j.jbiotec.2008.03.011

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 [微生物学]; 0836 [生物工程]; 090102 [作物遗传育种]; 100705 [微生物与生化药学];

摘要：

We have developed a novel method for typing single-nucleotide polymorphisms (SNPs) that can be applicable to rapid screening. The method involves the fusion of two PCR techniques, allele-specific PCR (AS-PCR) and L-DNA-tagged PCR (LT-PCR), which enables us to label PCR products with sequence-defined tags of mirror-image DNA (L-DNA). PCR products were applied without any purification or denaturation steps to gold surfaces where complementary single-stranded L-DNA was immobilized, and the products were detected with surface plasmon resonance (SPR) imaging. We were able to clearly discriminate 3 genotypes at position 2677 of the MDR1 gene (G/G-homozygote, G/T-heterozygote, and T/T-homozygote) by comparing SPR difference images. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：157 / 160

页数：4

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