Preparation of magnetite-loaded silica microspheres for solid-phase extraction of genomic DNA from soy-based foodstuffs.
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 microm 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.
Shi
Ruobing
R
School of Pharmaceutical Science & Technology, Tianjin University, Tianjin 300072, China.
Wang
Yucong
Y
Hu
Yunli
Y
Chen
Lei
L
Wan
Qian-Hong
QH
D000327:Adsorption; D002712:Chlorides; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005290:Ferric Compounds / Q000737:chemistry; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D030841:Food, Genetically Modified; D053768:Metal Nanoparticles / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D036103:Nanotechnology; D010316:Particle Size; D030821:Plants, Genetically Modified / Q000235:genetics; D016133:Polymerase Chain Reaction; D033362:Powder Diffraction; D012822:Silicon Dioxide / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods*; D045730:Soy Foods / Q000032:analysis*; D013025:Glycine max / Q000737:chemistry; D013053:Spectrophotometry; D013499:Surface Properties; D013698:Templates, Genetic
D002712:Chlorides; D005290:Ferric Compounds; D012822:Silicon Dioxide; D004247:DNA; C024555:ferric chloride; D052203:Ferrosoferric Oxide