Carles Maria C, Sucher Nikolaus J
Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA.
Methods Mol Biol. 2006;321:131-40. doi: 10.1385/1-59259-997-4:131.
The polymerase chain reaction (PCR) provides an in vitro method for rapid enzymatic amplification of fragments of DNA. Microchip-based PCR devices (with reaction volumes from picoliters to microliters) have been realized using various combinations of silicon, glass, and/or plastic materials. Passivation of exposed surfaces in the microreactor is critical for successful PCR. Silicon and plastic surfaces can be passivated by silanization. With surface passivation, PCR can be performed efficiently and economically in chip-based microreactors. The reduced thermal mass of microchips allows for extremely fast temperature ramping. PCR protocols established for benchtop reactors may need to be adjusted accordingly when transferred to microchips. Here, we provide detailed protocols for microchip PCR including procedures for surface passivation and bonding of glass to silicon with ultraviolet curable glue, because both procedures have a major influence on the success or failure of the PCR.
聚合酶链反应（PCR）提供了一种在体外快速酶促扩增DNA片段的方法。基于微芯片的PCR装置（反应体积从皮升至微升）已通过硅、玻璃和/或塑料材料的各种组合实现。微反应器中暴露表面的钝化对于成功进行PCR至关重要。硅和塑料表面可通过硅烷化进行钝化。通过表面钝化，可在基于芯片的微反应器中高效且经济地进行PCR。微芯片降低的热质量允许实现极快的温度变化。当将为台式反应器建立的PCR方案转移到微芯片上时，可能需要相应调整。在此，我们提供了微芯片PCR的详细方案，包括表面钝化以及用紫外光固化胶将玻璃与硅键合的程序，因为这两个程序对PCR的成败都有重大影响。
