Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, Virginia 22904, USA.
Analyst. 2010 Mar;135(3):531-7. doi: 10.1039/b918996c. Epub 2010 Jan 25.
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 beta-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 microL 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.
描述了一种新型的固相萃取技术,其中 DNA 以一种效率取决于磁珠的磁性操作而不是通过填充床的溶液流动的方式结合和洗脱。通过从全血中分离 DNA,随后扩增β珠蛋白基因的一个片段,证明了该技术从复杂样品中分离出相当纯净、可 PCR 扩增的 DNA 的实用性。通过在静态样品存在下有效地控制固相的运动,消除了在微通道中可重复地填充固相和保持一致流速的相关问题。这里描述的技术快速、简单、高效,允许从 0.6 μL 血液中回收超过 60%的 DNA,浓度适合 PCR 扩增。此外,这里提出的技术需要廉价的常见实验室设备,使其易于应用于临床即时护理应用和现场法医样本分析。