Suppr超能文献

利用磁泳技术对DNA进行连续微流纯化

Continuous Microfluidic Purification of DNA Using Magnetophoresis.

作者信息

Xu Ying, Zhang Zhen, Su Zhen, Zhou Xiaoxiang, Han Xiaoming, Liu Quanjun

机构信息

State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.

Shenzhen Research Institute of Southeast University, Shenzhen Virtual University Park, Shenzhen 518000, China.

出版信息

Micromachines (Basel). 2020 Feb 11;11(2):187. doi: 10.3390/mi11020187.

Abstract

Automatic microfluidic purification of nucleic acid is predictable to reduce the input of original samples and improve the throughput of library preparation for sequencing. Here, we propose a novel microfluidic system using an external NdFeB magnet to isolate DNA from the polymerase chain reaction (PCR) mixture. The DNA was purified and isolated when the DNA-carrying beads transported to the interface of multi-laminar flow under the influence of magnetic field. Prior to the DNA recovery experiments, COMSOL simulations were carried out to study the relationship between trajectory of beads and magnet positions as well as fluid velocities. Afterwards, the experiments to study the influence of varying velocities and input of samples on the DNA recovery were conducted. Compared to experimental results, the relative error of the final position of beads is less than 10%. The recovery efficiency decreases with increase of input or fluid velocity, and the maximum DNA recovery efficiency is 98.4% with input of l00 ng DNA at fluid velocity of 1.373 mm/s. The results show that simulations significantly reduce the time for parameter adjustment in experiments. In addition, this platform uses a basic two-layer chip to realize automatic DNA isolation without any other liquid switch value or magnet controller.

摘要

自动微流控核酸纯化技术有望减少原始样本的输入量,并提高测序文库制备的通量。在此,我们提出一种新型微流控系统,该系统使用外部钕铁硼磁体从聚合酶链反应(PCR)混合物中分离DNA。当携带DNA的磁珠在磁场影响下传输到多层流界面时,DNA被纯化并分离出来。在进行DNA回收实验之前,利用COMSOL软件进行模拟,以研究磁珠轨迹与磁体位置以及流体速度之间的关系。之后,开展了研究不同流速和样本输入量对DNA回收影响的实验。与实验结果相比,磁珠最终位置的相对误差小于10%。回收效率随样本输入量或流体速度的增加而降低,在流体速度为1.373 mm/s、输入100 ng DNA时,最大DNA回收效率为98.4%。结果表明,模拟显著减少了实验中参数调整的时间。此外,该平台使用基本的双层芯片实现了DNA的自动分离,无需任何其他液体开关阀或磁体控制器。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a6c/7074667/f04c68d7a925/micromachines-11-00187-g001.jpg

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验