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Characterization of dynamic solid phase DNA extraction from blood with magnetically controlled silica beads

机译:磁控硅胶珠从血液中动态固相DNA提取的表征

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摘要

A novel solid phase extraction technique is described where DNA is bound and eluted from magnetic silica beads in a manner where efficiency is dependent on the magnetic manipulation of the beads and not on the flow of solution through a packed bed. The utility of this technique in the isolation of reasonably pure, PCR-amplifiable DNA from complex samples is shown by isolating DNA from whole human blood, and subsequently amplifying a fragment of the b-globin gene. By effectively controlling the movement of the solid phase in the presence of a static sample, the issues associated with reproducibly packing a solid phase in a microchannel and maintaining consistent flow rates are eliminated. The technique described here is rapid, simple, and efficient, allowing for recovery of more than 60% of DNA from 0.6 mL of blood at a concentration which is suitable for PCR amplification. In addition, the technique presented here requires inexpensive, common laboratory equipment, making it easily adopted for both clinical point-of-care applications and on-site forensic sample analysis.
机译:描述了一种新颖的固相提取技术,其中以一种方式依赖于磁性二氧化硅珠子结合和洗脱DNA的方式,其效率取决于珠子的磁性操作,而不取决于溶液通过填充床的流量。通过从全血中分离DNA,然后扩增b-珠蛋白基因的片段,可以证明该技术在从复杂样品中分离合理纯度的,可PCR扩增的DNA中的实用性。通过有效地控制静态样品存在下固相的运动,消除了与可重复性地将固相填充到微通道中并保持恒定流速相关的问题。这里描述的技术是快速,简单和有效的,可从0.6 mL血液中以适合PCR扩增的浓度回收超过60%的DNA。此外,此处介绍的技术需要廉价的通用实验室设备,使其很容易用于临床即时护理应用和现场法医样品分析。

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