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玻璃微器件中的快速DNA扩增。

Rapid DNA amplification in glass microdevices.

作者信息

Easley Christopher J, Legendre Lindsay A, Landers James P, Ferrance Jerome P

机构信息

Department of Chemistry, University of Virginia, Charlottesville, VA, USA.

出版信息

Methods Mol Biol. 2006;339:217-32. doi: 10.1385/1-59745-076-6:217.

Abstract

The polymerase chain reaction (PCR) for amplification of DNA has become a very useful tool in scientific research and analytical laboratories, yet conventional techniques are time-consuming, and the reagents are expensive. Miniaturization of this technique has the potential to drastically reduce amplification time and reagent consumption while simultaneously improving the efficiency of the reaction. Increasing the surface area-to-volume ratio using microfluidic reaction chambers allows homogeneous solution temperatures to be achieved much more rapidly than in conventional heating blocks. Employing infrared radiation to selectively heat the reaction solution can additionally reduce the time and energy needed for thermocycling; the reaction container is not heated and can even serve as a heat sink for enhancement of cooling. Microchip systems also provide the potential for fabrication of structures for additional processing steps directly in line with the PCR chamber. Not only can amplification be integrated with product separation and analysis, but sample preparation steps can also be incorporated prior to amplification. The ultimate goal is a miniature total-analysis-system with seamlessly coupled sample-in/answer-out capabilities that consumes very low volumes of reagents and drastically reduces the time for analysis. This chapter will focus on the materials and methods involved in simple straight-channel microchip PCR on glass substrates using non-contact thermocycling.

摘要

用于DNA扩增的聚合酶链反应(PCR)已成为科研和分析实验室中非常有用的工具,但传统技术耗时且试剂昂贵。该技术的小型化有可能大幅缩短扩增时间并减少试剂消耗,同时提高反应效率。使用微流体反应腔增加表面积与体积之比,能比传统加热块更快地实现均匀的溶液温度。采用红外辐射选择性地加热反应溶液,还可减少热循环所需的时间和能量;反应容器不被加热,甚至可作为增强冷却的散热器。微芯片系统还为直接在与PCR腔室相连的线路上制造用于额外处理步骤的结构提供了可能性。不仅扩增可与产物分离和分析集成,而且样品制备步骤也可在扩增之前纳入。最终目标是一个微型全分析系统,具有无缝耦合的进样/出结果能力,消耗极低体积的试剂并大幅缩短分析时间。本章将重点介绍在玻璃基板上使用非接触热循环进行简单直通道微芯片PCR所涉及的材料和方法。

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