基于纳米水凝胶颗粒的毛细管电泳法分离DNA  

Separation of DNA by Capillary Electrophoresis Based on Nanoparticles Hydrogel

作　　者：张禾蓉 易达 孟子晖 薛敏 汪海林[2] 冯金生 ZHANG He-Rong;YI Da;MENG Zi-Hui;XUE Min;WANG Hai-Lin;FENG Jin-Sheng(School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 102488, China;Research Center For Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China)

机构地区：[1]北京理工大学化学与化工学院,北京102488 [2]中国科学院生态环境研究中心,北京100085

出　　处：《分析化学》2019年第5期772-778,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.21874009)资助~~

摘　　要：采用乳液聚合法,在水相中制备了一系列纳米水凝胶颗粒(NPs),基于动态涂层法应用于毛细管电泳分离DNA片段。动态光散射表征结果表明,所制备的NPs分散性好、粒径分布均匀。基于NPs交联网络结构将其作为毛细管电泳分离介质,结果表明,单体的用量、纳米颗粒的粒径以及NPs在毛细管电泳中的浓度对DNA分离效果都有影响。当单体采用N-异丙基丙烯酰胺(87%)、丙烯酸羟乙酯(8%)和丙烯酸(4%),交联剂为N,N'-亚甲基双丙烯酰胺(1%)时,通过改变表面活性剂十二烷基磺酸钠(SDS)的用量,可以改变所制备的纳米颗粒粒径,当SDS用量为53 mg时,NPs粒径约480 nm。将此NPs(8.0 mg/mL)加入TG缓冲溶液(25 mmol/L Tris,192 mmol/L甘氨酸,pH 8.3)用于毛细管电泳,3个长度DNA片段(20、50和80 nt)可实现完全分离,分离效果好,且迁移时间较短。A library of Nanoparticles hydrogel (NPs) was synthesized by emulsion polymerization method in aqueous solution. The feasibility of NPs applying to the DNA fragments separation in capillary electrophoresis was investigated based on the dynamic coating method. The diameters of the particles were proved uniform with good mono-dispersion by dynamic light scattering method. The experimental results indicated that three factors had an effect on the DNA separation, including the amount of monomer, the particle size and the concentration of NPs in capillary electrophoresis buffer solution. NPs exhibited the best performance in DNA separation when the monomer molar ratios were optimized to N -isopropylacrylamide (87%), hydroxyethylacrylate (8%), acrylic acid (4%) and N,N '-methylene bisacrylamide(1%). Meanwhile, the particle size of the prepared NPs could be changed by changing the amount of the surfactant sodium dodecyl sulfate (SDS). A completely separation among the three different length of DNA frament samples (20 nt, 50 nt and 80 nt) could be obtained when 8.0 mg/mL NPs (400 nm) were added in the TG buffer solution ( 25 mmol/L Tris, 192 mmol/L glycine, at pH 8.3) in the capillary electrophoresis study. These hydrogel nanoparticles showed potential capability in DNA separation due to their well-developed crosslinking network structure.

关 键 词：纳米水凝胶颗粒 毛细管电泳 DNA分离 

分 类 号：O658.9[理学—分析化学] Q503[理学—化学]