High-throughput SNP detection using nano-scale engineered biomagnetite
High-throughput SNP detection using nano-scale engineered biomagnetite
复制标题
DOI：
10.1016/j.bios.2006.12.022
复制
发表时间：
2007-04-15
影响因子：
12.6
通讯作者：
Katoh, Takahiko
中科院分区：
文献类型：
--
作者：
Matsunaga, Tadashi;Maruyama, Kohei;Katoh, Takahiko
基于等位基因特异性寡核苷酸、杂交和热解离曲线分析，使用纳米级工程生物磁铁矿（细菌磁性颗粒; BacMP）开发了用于大规模检测单核苷酸多态性（SNP）的半自动化系统。为了在大量样品中进行可靠的检测，优化了在纳米尺寸的BacMP上捕获靶DNA和双链DNA变性的几种条件。使用短PCR扩增子（69 bp）观察到最有效的靶DNA捕获。使用50 mM NaOH使捕获的DNA变性。通过这些优化，对822个转化生长因子（TGF）-β 1基因样本进行了大规模SNP检测，该基因富含GC含量和重复序列。与传统的基于测序的方法相比，证实了基于BacMP的半自动SNP检测系统的高可靠性。(c)2007 Elsevier B. V.保留所有权利。
A semi-automated system for the large-scale detection of single nucleotide polymorphisms (SNPs) has been developed based on allele-specific oligonucleotide, hybridization and thermal dissociation curve analysis using nano-scale engineered biomagnetite (bacterial magnetic particles; BacMPs). For reliable detection in large numbers of samples, several conditions for the capture of target DNA on nano-sized BacMPs and the denaturation of double-stranded DNA were optimized. The most efficient target DNA capture was observed using short PCR amplicons (69 bp). Captured DNAs were denatured using 50 mM NaOH. With these optimizations, large-scale SNP detection was performed on 822 samples of the transforming growth factor (TGF)-beta 1 gene, which is rich in both GC content and repetitive sequences. High reliability for the semi-automated BacMP-based SNP detection system was confirmed following comparison to traditional sequencing-based methods. (c) 2007 Elsevier B.V. All rights reserved.