Non-contact microchip PCR based on infrared Heating

描述(申请人提供)：尽管目前流行‘基因组时代’即将结束(随着‘后基因组时代’的开始)，基因分析无疑将继续是一个多产的探索领域。遗传分析方法对于充实与肿瘤生长相关的新突变、了解单核苷酸多态的意义以及开发更好的微卫星DNA基因分型方法将是至关重要的。因此，从粗制生物样品中提纯核酸物质并通过聚合酶链式反应(PCR)成功扩增目标DNA序列仍将是一项至关重要的必要程序。我们已经证明了在微芯片中使用IR中介加热(即使用加热灯)执行PCR的可行性，并表明这可以直接与同一微芯片上的分离相结合。我们现在建议通过定义适当的微芯片设计和相应的仪器来促进这些发展，以便在单个微芯片上进行多路聚合酶链式反应(一次36个样本)和DNA分离。PCR扩增将是完全非接触的(红外加热和干涉测温)，能够在15分钟或更短的时间内完成DNA扩增(最多25个循环)。随后将在同一微芯片上对聚合酶链式反应产物进行多通道电泳分析，样品吞吐量超过每分钟一个样品(在不到30分钟内有32个样品)。多路聚合酶链式反应与微设备上分离的无缝集成将向自动化仪器迈进一大步，该仪器具有用于诊断和各种其他领域的“样本输入-回答”能力。这种方法不仅将利用微芯片技术的力量来减少样本量和提高聚合酶链式反应过程的速度，而且还将为在微芯片上进行真正的集成基因分析(也就是真正的芯片实验室)奠定基础。

DESCRIPTION (provided by applicant): Despite the current fad that the 'genome era' is coming to an end (as the 'post-genome era' gets underway), genetic analysis will undoubtedly continue to be a prolific field of exploration. Genetic analytical methods will be critical in fleshing out new mutations correlative of neoplastic growth, understanding the significance of single nucleotide polymorphisms, and developing better genotyping methods for, e.g., microsatellite DNA. Accordingly, the purification of nucleic acid material from crude biological samples and the successful amplification of target DNA sequences by the polymerase chain reaction (PCR) will remain a requisite protocol that is of vital importance. We have demonstrated the feasibility of executing PCR in microchips using IR-mediated heating (i.e., using a heat lamp) and have shown that this can be directly interfaced with separation on the same microchip. We now propose to further these developments by defining the appropriate microchip design and corresponding instrumentation for multiplexed PCR (36 samples at a time) and DNA separation on a single microchip. The PCR amplification will be completely non-contact (IR for heating and interferometry for temperature sensing), and capable of completing DNA amplification (up to 25cycles) in 15 minutes or less. Multi-channel electrophoretic analysis of the PCR products will ensue on the same microchip with a sample throughput of more than a sample per minute (32 samples in less than 30 minutes). The seamless integration of multiplexed PCR with separation on a microdevice will provide a giant step towards automated instrumentation with 'sample-in-answer-out' capabilities for diagnostics and a variety of other fields. Such an approach will not only exploit the power of microchip technology for reducing sample volumes and increasing the speed of the PCR process, but set the stage for truly integrated genetic analysis on a microchip (a.k.a., a true lab-on-a-chip).