Micro-Solid Phase Extraction of DNA on Microchips

描述（由申请人提供）： 尽管目前流行“后基因组”时代即将结束，但遗传分析很可能将继续成为一个多产的探索领域。遗传分析方法将是至关重要的充实新的突变相关的肿瘤生长，了解单核苷酸多态性的意义，并开发更好的微卫星DNA基因分型方法。因此，从粗制生物样品中纯化核酸材料，这是通过聚合酶链式反应（PCR）成功扩增靶DNA序列的必要方案，仍然是至关重要的。开发生物样品（全血、细胞培养物等）中DNA的固相萃取（SPE）方案在上一个资助期内，已证明以微型芯片形式制作是可行的。该平台利用微芯片技术的力量来减少样品体积并提高提取过程的速度，但需要进行优化以最大限度地发挥该方法的潜力。微芯片平台的另一个好处是能够集成额外的处理步骤。这将通过开发一种基于SPE的微芯片DNA纯化方案来研究，该方案与芯片上PCR直接无缝匹配，为微芯片上真正集成的遗传分析奠定基础（又名，真正的芯片实验室）。开发工作的主要部分还将集中在SPE和PCR步骤之间直接在微芯片上集成DNA定量方法。这将允许在PCR扩增中最佳地使用从样品中提取的DNA。

DESCRIPTION (provided by applicant): Despite the current fad that the 'post-genome' era is coming to an end, it is likely that genetic analysis will 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 microsatellite DNA. Accordingly, the purification of nucleic acid material from crude biological samples, a requisite protocol for successful amplification of target DNA sequences by the polymerase chain reaction (PCR), is still of vital importance. Development of a protocol for solid phase extraction (SPE) of DNA from biological samples (whole blood, cell culture, etc.) has been shown to be feasible in a microchip format during the previous funding period. This platform exploits the power of microchip technology for reducing sample volumes and increasing the speed of the extraction process, but optimization is needed to maximize the potential of this method. The additional benefit of the microchip platform is the ability to integrate additional processing steps. This will be investigated through the development of a microchip SPE-based DNA purification protocol that is directly and seamlessly mated with on-chip PCR, setting the stage for truly integrated genetic analysis on a microchip (a.k.a., a true lab-on-a-chip). A major part of the development work will also focus on integration of DNA quantitation methods directly on the microchip between the SPE and PCR steps. This will allow optimal use of the DNA extracted from the sample in the PCR amplification.