Preparation of magnetite-loaded silica microspheres for solid-phase extraction of genomic DNA from soy-based foodstuffs
Preparation of magnetite-loaded silica microspheres for solid-phase extraction of genomic DNA from soy-based foodstuffs
复制标题
负载磁铁矿的二氧化硅微球的制备用于固相提取大豆食品中的基因组 DNA
DOI：
10.1016/j.chroma.2009.07.016
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发表时间：
2009-09-04
影响因子：
4.1
通讯作者：
Wan, Qian-Hong
中科院分区：
文献类型：
--
作者：
Shi, Ruobing;Wang, Yucong;Wan, Qian-Hong
固相萃取已广泛用于制备用于基于聚合酶链反应（PCR）的分析方法的DNA模板。在研究的各种吸附剂中，磁响应二氧化硅颗粒由于其简化、加速和自动化提取过程的潜力而特别有吸引力。在这里，我们报告了一个简单的方法制备这样的磁性颗粒，这需要浸渍的多孔二氧化硅微球与铁盐，然后煅烧和还原处理。采用粉末X射线衍射仪（XRD）、扫描电子显微镜（SEM）、氮气吸附/脱附等温线和振动样品磁强计（VSM）对样品进行了表征。XRD数据显示，还原后在二氧化硅载体的孔道内产生约27.2nm的磁铁矿纳米晶体。SEM图像显示，合成的颗粒呈现球形形状和约3 μ m的均匀粒度，由二氧化硅载体确定。氮吸附数据证实，磁铁矿负载的二氧化硅颗粒具有典型的介孔结构，BET表面积约为183 m2/g。VSM数据表明，该颗粒具有顺磁性，饱和磁化强度为11.37emu/g。以磁性二氧化硅微球为载体，研究了其对大豆基因组DNA的快速提取效果。通过PCR扩增纯化的DNA模板用于筛选转基因生物（GMOs）。初步结果证实，使用磁铁矿负载的二氧化硅微球的DNA提取方案能够产生DNA模板，其是无杂环的，并准备用于下游分析。(C)2009爱思唯尔有限公司版权所有。
Solid-phase extraction has been widely employed for the preparation of DNA templates for polymerase chain reaction (PCR)-based analytical methods. Among the variety of adsorbents studied, magnetically responsive silica particles are particularly attractive due to their potential to simplify, expedite, and automate the extraction process. Here we report a facile method for the preparation of such magnetic particles, which entails impregnation of porous silica microspheres with iron salts, followed by calcination and reduction treatments. The samples were characterized using powder X-ray diffractometry (XRD), scanning electron microscopy (SEM), nitrogen adsorption/desorption isotherms, and vibrating sample magnetometry (VSM). XRD data show that magnetite nanocrystals of about 27.2 nm are produced within the pore channels of the silica support after reduction. SEM images show that the as-synthesized particles exhibit spherical shape and uniform particle size of about 3 mu m as determined by the silica support. Nitrogen sorption data confirm that the magnetite-loaded silica particles possess typical mesopore structure with BET surface area of about 183 m(2)/g. VSM data show that the particles display paramagnetic behavior with saturation magnetization of 11.37 emu/g. The magnetic silica microspheres coated with silica shells were tested as adsorbents for rapid extraction of genomic DNA from soybean-derived products. The purified DNA templates were amplified by PCR for screening of genetically modified organisms (GMOs). The preliminary results confirm that the DNA extraction protocols using magnetite-loaded silica microspheres are capable of producing DNA templates which are inhibitor-free and ready for downstream analysis. (C) 2009 Elsevier B.V. All rights reserved.