NON CONTACT INFRARED MEDIATED THERMOCYCLING FOR CHIP PCR

描述（改编自研究者摘要）： 对构成DNA的约3 X 10 exp 9碱基对进行测序， 人类基因组是人类基因组中承担的巨大任务 倡议（HGI）。 虽然分子生物学技术的进步使 这可能是测序方案中的方法学“瓶颈”， 由于必须通过以下方式分离测序片段， 常规平板凝胶电泳。 因此， 在增强传统的平板凝胶为基础的方法与新的兴趣 可以提高测序通量的技术。 两个相对较新的 电泳形式，毛细管电泳和电泳在 微制造的“芯片”已经显示出提供这种技术的前景。 尽管存在一些固有的技术挑战， 蚀刻在平面玻璃表面的多个通道中的分离 或塑料基底可以显著提高测序速度。 然而，在这方面， 芯片平台的真正力量不仅来自于 快速的电泳分离，但从能力无缝 将样品制备集成到芯片中。 这将允许直接和 PCR和电泳在单个设备中的自动化接口。 这项建议的目的是开发一种新的热循环方法 与其他方法不同的是，它可以很容易地集成到芯片中， 平台 PCR将使用基于红外的热循环进行 利用钨丝灯作为简单但有效的加热器的技术 输送系统，结合红外检测/控制溶液 温度 结合起来，这将允许PCR在芯片上执行 而不接触芯片。 因此，成本- PCR的时间效率将大大提高， 方法论 这一发展将为快速连接 循环测序反应与先决条件的样品净化过程， PCR后电泳分离，全部在一个单一的一次性 微量分析装置

DESCRIPTION (Adapted from the Investigator's Abstract): The mapping and sequencing of the approximately 3 X 10exp9 base pairs of DNA that constitute the human genome is the colossal task undertaken in the Human Genome Initiative (HGI). While advances in molecular biological techniques make this possible, a methodological "bottleneck" in the sequencing protocol has been created by the necessity to separate sequencing fragments via conventional slab gel electrophoresis. Consequently, there has been intense interest in augmenting the conventional slab gel-based methodology with new technologies that can enhance sequencing throughput. Two relatively new electrophoretic formats, capillary electrophoresis and electrophoresis in microfabricated "chips," have shown promise for providing this technology. Despite some inherent technical challenges, the ability to execute ultrafast separations in multiple channels etched into the surface of a planar glass or plastic substrate could enhance sequencing speed significantly. However, the true power of the chip platform stems, not only from the capability for rapid electrophoretic separations, but from the ability to seamlessly integrate sample preparation into the chip. This would allow for direct and automated interfacing of PCR and electrophoresis in a single device. The object of this proposal is to develop a novel thermocycling approach that, unlike other approaches, can easily be integrated into the chip platform. PCR will be carried out using an infrared-based thermocycling technology that exploits a tungsten lamp as a simple but effective heat delivery system, coupled with infrared detection/control of solution temperature. Combined, this will allow for PCR to be executed on the chip with nanoliter volumes without contacting the chip. Consequently, the cost- and time-efficiency of PCR will be improved dramatically over conventional methodology. This development will set the stage for interfacing rapid cycle sequencing reactions with the prerequisite sample clean-up process and the post-PCR electrophoretic separation, all in a single, disposable microanalysis device.