基于微通道多向阀自动化快速提取DNA的研究  

Automated Rapid DNA Extraction Based on Microchannel Multidirectional Valves

作　　者：高英俊 付立申 徐晓龙 吴文明 GAO Ying-jun;FU Li-shen;XU Xiao-long;WU Wen-ming(School of Environmental and Chemical Engineering,Wuyi University,Jiangmen 529020,China;Institute of Biological and Medical Engineering,Guangdong Academy of Sciences,Guangzhou 510316,China;School of Applied Physics and Materials,Wuyi University,Jiangmen 529020,China)

机构地区：[1]五邑大学环境与化学工程学院,广东江门529020 [2]广东省科学院生物与医学工程研究所,广东广州510316 [3]五邑大学应用物理与材料学院,广东江门529020

出　　处：《五邑大学学报（自然科学版）》2024年第3期7-12,共6页Journal of Wuyi University(Natural Science Edition)

基　　金：广东省科学院发展专项资金项目(2022GDASZH-2022010110,2023GDASZH-2023010102)。

摘　　要：核酸的高效分离与纯化对于下游分子诊断具有至关重要的影响.本研究基于磁珠法提取原理设计出快速、自动化核酸提取系统,利用步进电机和注射泵组成的移液装置通过微通道连接多向电磁阀完成核酸裂解、洗涤、洗脱、纯化,通过大鼠主动脉内皮细胞进行核酸提取和扩增,对核酸加热裂解时间进行优化.结果表明,当裂解时间大于4 min时,核酸纯化效果最好,磁珠回收率可达96.5%,紫外吸收光比值大于1.8,同时整个自动化提取仅需8 min.通过微通道多向阀与核酸提取技术纯化后的核酸可直接用于扩增实验,且与手动提取相比聚合酶链式反应的Ct值起跳更早,有利于后续集成核酸检测一体化系统.Efficient isolation and purification of nucleic acids play a crucial role in downstream molecular diagnostics.In this study,a rapid and automated nucleic acid extraction system was designed based on the principle of magnetic bead extraction.A pipette consisting of a stepper motor and an injection pump was used to complete nucleic acid lysis,washing,elution and purification by connecting a multi-directional solenoid valve through a microchannel,and the nucleic acid was extracted and amplified by a rat’s aortic endothelial cells,and nucleic acid heating and lysis time was optimized.Results showed that when the cracking time exceeded 4 minutes,the nucleic acid purification effect was optimal,with a magnetic bead recovery rate of up to 96.5%and a UV absorbance ratio exceeding 1.8.The entire automated extraction process required only 8 minutes.Nucleic acids purified by microchannel multi-way valve and nucleic acid extraction technology can be directly used for amplification experiments,and the Ct value of PCR can commence earlier than that of manual extraction,which is conducive to the subsequent integration of nucleic acid detection integration system.

关 键 词：样品预处理 核酸提取 微通道 磁珠法 

分 类 号：R331[医药卫生—人体生理学]