Angew Chem Int Ed:清華大學劉東生研究組發(fā)展出基于酸堿調控的室溫PCR方法

近日，《Angew. Chem. Int. Ed.》雜志上發(fā)表了中國科學院上海應用物理研究所樊春海課題組與清華大學陸東生研究組合作的一篇研究論文，論文報道了一種新型的離子介導聚合酶鏈式反應(Ion-Mediated Polymerase Chain Reaction, IM-PCR)，通過精確調控溶液pH值(即質子和氫氧根離子)，可以在室溫下完成PCR擴增。
PCR是體外通過酶反應合成、高靈敏擴增目標基因片段的一種方法，是常用的分子生物學技術之一。常規(guī)PCR是通過控制溶液溫度來實現DNA分子的可逆變復性，從而達到復制擴增的目的。而這種升降溫過程通常需要復雜的控溫裝置并消耗大量能源。從理論上來說，DNA變復性還可以通過酸堿變性來實現，然而如何在不改變溶液體積的情況下精確控制溶液pH值則是一個難題。
研究人員建立了一種基于微流控的電化學芯片，可以通過電壓驅動快速、精確調控溶液pH值，并實現了對DNA分子機器的電驅動控制(Nano Lett. 2010,10,1393)?；谶@一研究基礎，上海應物所博士張一和副研究員李茜發(fā)現微流控電化學可以有效控制PCR反應體系的pH值，實現了室溫下的酸堿驅動的IM-pcr擴增。與傳統(tǒng)PCR技術相比，這一新型的集成化IM-PCR技術無需變溫過程，所有反應步驟均在室溫下進行。這一小型化、低成本、易于集成的PCR新技術有望在生物檢測、臨床診斷及環(huán)境監(jiān)測中發(fā)揮作用。
Angew Chem Int Ed:清華大學劉東生研究組發(fā)展出基于酸堿調控的室溫PCR方法
基于酸堿調控的室溫PCR新方法
原文鏈接：
Ion-Mediated Polymerase Chain Reactions Performed with an Electronically Driven Microfluidic DevICE

原文摘要：
The polymerase chain reaction (PCR) is a powerful method for exponentially amplifying very low amounts of target DNA from genetic, clinical, and forensic samples. However, the heating and cooling steps in PCR largely hamper the miniaturization of thermocyclers for on-site detection of pathogens and point-of-care tests. Herein, we devise an ion-mediated PCR (IM-PCR) strategy by exploiting ion-induced DNA denaturation/renaturation cycles. DNA duplexes are effectively denatured in alkaline solutions; whereas, the denatured single-stranded DNA strands readily reform duplexes at neutral pH. By using an integrated microchip that can programmably control the solution pH simply switching the potential in a range of several hundred millivolts, we can trigger IM-PCR at a constant temperature. Analogously to thermal cycling, 30 cycles of pH-induced denaturation/renaturation were used to amplify protein DNA fragments as confirmed by DNA sequencing. We anticipate that this portable, low-cost, and scalable IM-PCR holds great promise for widespread biological, clinical, and environmental applications.