摘要
制备了表面电性可控的氨基化SiO_2@Fe_3O_4磁性复合微球,采用激光刻蚀和热压键合的方法制作了聚甲基丙烯酸甲酯(PMMA)材质的DNA固相萃取芯片,将磁珠灌注于芯片通道中,借助永磁铁固定并控制磁珠,将磁珠芯片应用于人类全血中的基因组DNA提取,优化了提取实验条件,并对提取产物进行凝胶电泳和PCR分析。实验结果表明,磁珠微流控芯片成功地从全血中提取出纯度较高的基因组DNA,提取效率约35%,提取液的凝胶电泳条带与商品化试剂盒提取的基因组DNA一致,提取液可用于进一步的PCR反应。
Self-made surface charge switching amino-SiO 2@Fe 3O 4 magnetic composite microspheres were applied to microfluidic chip,and DNA solid phase extraction chip of polymethyl methacrylate(PMMA)was fabricated by laser etching and hot pressing bonding.The magnetic beads that could be controlled by permanent magnet were injected into the chip channel to obtain a magnetic bead chip for genomic DNA extraction in human whole blood.Under the optimized extraction conditions(with 80μg beads,flow rate for adsorption,elution and desorption were 10,5 and 20μL/min,respectively),the extracted products were subjected to gel electrophoresis and PCR test.The results showed that the genomic DNA with high purity was extracted from the whole blood by magnetic bead microfluidic chip.The extraction efficiency was about 35%.The gel electrophoresis band of the extract was consistent with the genomic DNA extracted by commercial reagent kit.The extract solution could be used for further PCR reaction.
作者
黄华斌
傅奇
胡佳
游其华
庄峙厦
杨朝勇
王小如
HUANG Hua-Bin;FU Qi;HU Jia;YOU Qi-Hua;ZHUANG Zhi-Xia;YANG Chao-Yong;WANG Xiao-Ru(XiaMen HuaXia College,Department of Science and Technology for Inspection,Xiamen 361024,China;College of Chemistry and Chemical Engineering,Xiamen University,Xiamen 361005,China)
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2018年第9期1372-1378,共7页
Chinese Journal of Analytical Chemistry
基金
国家星火计划项目(No.2015GA721002)
黄土与第四纪地质国家重点实验室开放基金(No.SKLLQG1607)
福建省中青年教师科技计划项目(No.JAT170817)资助~~
关键词
表面电性可控
磁珠
微流控芯片
DNA提取
Surface charge switching
Magnetic beads
Microfluidic chip
DNA extraction
作者简介
王小如,E-mail:wxr@hxxy.edu.cn。
