Fabrication and application of single nucleotide polymorphisms library on magnetic nanoparticles using adaptor PCR
Fabrication and application of single nucleotide polymorphisms library on magnetic nanoparticles using adaptor PCR
复制标题
适配器PCR磁性纳米粒子单核苷酸多态性文库的构建及应用
DOI：
10.1166/jnn.2008.014
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发表时间：
2008-01-01
影响因子：
--
通讯作者：
He, Nongyue
中科院分区：
文献类型：
--
作者：
Liu, Hongna;Li, Song;He, Nongyue
我们开发了一种基于衔接子PCR的磁性纳米颗粒（MNP）单核苷酸多态性（SNPs）文库构建的新方法。通过与一对等位基因特异性双色荧光（Cy 3，Cy 5）探针杂交来询问文库中的每个SNP位点以确定SNP。应用该方法检测血管紧张素原（AGT）基因中与原发性高血压相关的两个SNPs位点（M235 T和A-6 G），并对其荧光信号进行定量。纯合基因型的荧光比率（匹配探针：错配探针信号）超过3.0，而杂合基因型的比率接近1.0。该方法不需要复杂的多重PCR过程，可以在有限的个体DNA样本中进行SNPs的多重检测，是一种简单、高效、可靠的方法。
We have developed a novel approach to fabricate single nucleotide polymorphisms (SNPs) library on magnetic nanoparticles (MNPs) based on adaptor PCR. Each SNP locus in the library was interrogated by hybridization with a pair of allele specific dual-color fluorescence (Cy3, Cy5) probes to determine SNP. Two SNPs loci (M235T and A-6G) associated with essential hypertension in the angiotensinogen (AGT) gene were detected by this method and their fluorescent signals were quantified. The fluorescent ratios (match probe: mismatch probe signal) of homozygous genotypes were over 3.0, whereas heterozygous genotypes had ratios near to 1.0. Without any complex multiplex PCR procedure, it is a simple, efficient and reliable method for the multiplex SNPs detection using limited amount of DNA samples from individuals.