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Dielectrophoretic Manipulation of DNA: Separation and Polarizability

机译:DNA的介电泳操作:分离和极化率。

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Although separation of polymers based on the combinationof dielectrophoretic trapping and electrophoretic forces was proposed 15 years ago, experimental proof has not yet been reported. Here, we address this problem for long DNA fragments in a simple and easy-to-fabricate microfluidic device, in which the DNA is manipulated by electrophoresis and by electrodeless dielectrophoresis. By slowly increasing the strength of the dielectrophoretic traps in the course of the separation experiments, we are able to perform efficient and fast DNA separation according to length for two different DNA conformations: linear DNA (lambda (48.5-kbp) and T2 (164-kbp) DNA) and supercoiled covalently closed circular plasmid DNA (7 and 14 kbp). The underlying migration mechanism-thermally induced escape processes out of the dielectrophoretic traps in the direction of the electrophoretic force-is sensitive to different DNA fragments because of length-dependent DNA polarizabilities. This is analyzed in a second series of experiments, where the migration mechanism is exploited for the quantitative measurement of the DNA polarizabilities. This new and simple technique allows for the systematic characterization of the polarizability not only for DNA but also for other biomolecules like proteins. Furthermore, our results have direct implications to future biotechnological applications such as gene therapy and DNA vaccination.
机译:尽管在15年前就提出了基于介电电泳和电泳力的聚合物分离方法,但尚未有实验证明。在这里,我们通过简单且易于制造的微流体装置解决了长DNA片段的问题,在该装置中,DNA通过电泳和无电极介电电泳操作。通过在分离实验过程中缓慢增加介电泳陷阱的强度,我们能够根据长度对两种不同的DNA构象进行有效而快速的DNA分离:线性DNA(λ(48.5-kbp)和T2(164- kbp)DNA)和超螺旋共价封闭的环状质粒DNA(7和14 kbp)。潜在的迁移机制-在电泳力的方向上从介电泳陷阱中热诱导的逃逸过程-由于长度依赖的DNA极化能力,对不同的DNA片段敏感。在第二系列实验中对此进行了分析,其中利用了迁移机制来定量测量DNA极化率。这种新的简单技术不仅可以对DNA进行极化,而且还可以对蛋白质等其他生物分子进行极化特性的系统表征。此外,我们的结果对未来的生物技术应用(如基因治疗和DNA疫苗接种)具有直接影响。

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