硅磁微球的制备及其在植物核酸分离纯化中的应用  

Preparation of Silicon Magnetic Microspheres and Its Application in Nucleic Acid Extraction

作　　者：刘涛[1] 张宇[2] 王丽娜[1] LIU Tao;ZHANG Yu;WANG Li-na(State Key Laboratory of Forest Genetics and Breeding,Northeast Forestry University,Harbin 150040,China;.College of Law/Harbin University of Commerce,Harbin 150028,China)

机构地区：[1]东北林业大学林木遗传育种国家重点实验室,黑龙江哈尔滨150040 [2]哈尔滨商业大学法学院,黑龙江哈尔滨150028

出　　处：《山东农业大学学报（自然科学版）》2023年第3期378-384,共7页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然基金青年项目:杨树PtPEPCK1调控碳氮平衡的分子机理研究(31800565)。

摘　　要：本文采用改进的共沉淀方法制备Fe3O4纳米粒子,并利用表面活性剂柠檬酸对其表面改性;然后利用溶胶凝胶法,催化TEOS水解和缩合反应,使SiO2包裹在Fe3O4纳米粒子表面,成功制备出了Fe3O4@SiO2复合纳米粒子。结果表明,Fe3O4@SiO2复合纳米粒子粒径在200-400 nm之间,粒径均匀,具有超顺磁性,磁响应时间在10-20 s,半沉降时间在20 min。根据高盐吸附低盐洗脱原理,建立了一种以磁性微球为载体的核酸快速提取和纯化方法。在白桦基因组DNA提取中,OD260/OD280均在1.8左右,OD260/OD230均在2.0以上,提取的质量和浓度比传统方法要好。在白桦RNA提取过程中,OD260/OD280和OD260/OD230均在2.0以上,提取的RNA浓度要比传统方法更高。磁珠法与传统方法比较时,核酸提取纯化效果上更佳,提取速度快、安全、成本低,具有很高的推广价值。Fe3O4@SiO2 composite nanoparticles were successfully prepared by using a modified co-precipitation method to prepare Fe3O4 nanoparticles and using the surfactant citric acid to modify their surface;then using the sol-gel method to catalyze the TEOS hydrolysis and condensation reaction to wrap SiO2 on the surface of Fe3O4 nanoparticles.The results showed that the Fe3O4@SiO2 composite nanoparticles were in the range of 200-400 nm with uniform particle size,superparamagnetic,magnetic response time in the range of 10-20 s,and semi-sedimentation time in the range of 20 min.Based on the principle of high salt adsorption and low salt elution,a rapid extraction and purification method of nucleic acids using magnetic microspheres as carriers was established.In the extraction of Betula platyphylla genomic DNA,OD260/OD280 were all around 1.8 and OD260/OD230 were all above 2.0,and the quality and concentration of extraction were better than the traditional method.In the Betula platyphylla RNA extraction,OD260/OD280 and OD260/OD230 were both above 2.0,and the extracted RNA concentration was higher than that of the traditional method.When the magnetic bead method was compared with the traditional method,the nucleic acid extraction and purification was better,and the extraction was fast,safe and low cost,which has high promotion value.

关 键 词：硅磁微球 植物核酸 分离纯化 

分 类 号：Q943.2[生物学—植物学] TB383[一般工业技术—材料科学与工程]