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，但基因分析很可能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、了解单核苷酸多态性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，从粗制的生物样品中提纯核酸物质，作为通过聚合酶链式反应(PCR)成功扩增目标DNA序列的必要程序，仍然是至关重要的。生物样品(全血、细胞培养等)中DNA固相萃取(SPE)方法的建立在上一个供资期间，微芯片格式已被证明是可行的。该平台利用了微芯片技术的力量，减少了样品体积，提高了提取过程的速度，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微芯片SPE的DNA纯化协议来进行研究，该协议直接与芯片上的PCR无缝配对，为在微芯片上进行真正的集成基因分析奠定基础(也就是真正的芯片实验室)。开发工作的一个主要部分还将侧重于将DNA定量方法直接整合到SPE和PCR步骤之间的微芯片上。这将使从样品中提取的DNA在聚合酶链式反应扩增中得到最佳使用。

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.