PIEZO ELECTRIC PIPETTING TECHNOLOGY FOR DNA ANALYSIS

这项资助结合了两项创新技术：一种新的流体传输设备和一种在cDNA阵列上检测单核苷酸多态(SNPs)的新技术。这两项技术结合在一起，将对癌症的分子分析产生立竿见影的重大影响。在压电式元件的驱动下，流体传输技术能够实现亚纳升体积的流体的移液。此外，新的传感技术允许持续监测设备的运行状态。传感技术对于大规模并行DNA分析应用是必不可少的，这些应用需要可靠地吸入和分配数千个不同的样本。最终目标是将流体传输和传感器技术集成到用于癌症研究和一般实验室应用的自动移液仪器中。最初，该仪器将应用于一项新技术，使其首次有可能在现成且相对便宜的斑点cDNA阵列上检测SNPs、缺失和插入。这位合作研究人员已经向华盛顿州西雅图的弗雷德·哈钦森癌症研究中心(F.H.C.R.C.)提交了一份关于这项令人兴奋的新技术的发明披露。到目前为止，检测DNA微阵列上的SNPs只能使用Affymetrix生产的寡核苷酸阵列(GeneChips)上的等位基因特异性杂交。这些基因芯片的有限获取及其高昂的成本将使这项技术难以应用于大量DNA样本。拟议研究的一个主要目标是开发和论证新的SNP检测方法的可行性，该方法使用将在本拨款中开发的新型阵列产生的高密度cDNA阵列来高通量地检测多个基因座的多态。这些高重复性、高密度的阵列也有望在分析mRNA表达时提供更高的灵敏度和特异度。将进行实验，以证明所提出的技术相对于现有的基于PIN的机械阵列在基因表达分析中的优势。拟议的商业应用：在生物芯片、微阵列和高通量筛选等新兴技术以及普遍高昂的试剂成本的推动下，生物医疗行业正朝着使用较小流体样本量的分析方向发展。这项拟议的研究直接针对这一不断增长的市场，为这些应用开发了一种通用的自动化实验室移液仪器。通过降低开发成本，这项新技术将使更广泛的癌症研究界能够获得高质量、高密度的DNA阵列。

This grant brings together two innovative technologies: a novel fluid transfer device and a novel technique for detecting Single Nucleotide Polymorphisms (SNPs) on cDNA arrays. Integrated together, these two technologies will have an immediate and significant impact on the molecular analysis of cancer. Actuated by piezo-electric elements, the fluid transfer technology enables the pipetting of sub-nanoliter volumes of fluid. Additionally, novel sensing technology allows the operational state of the device to be continuously monitored. The sensing technology is indispensable for massively parallel DNA analysis applications requiring thousands of different samples to be reliably aspirated and dispensed. The ultimate goal is to integrate the fluid transfer and sensor technology into an automated pipetting instrument for cancer research and general laboratory applications. Initially the instrument will be applied to a novel technology making it possible for the first time to detect SNPs, deletions and insertions on readily available and relatively inexpensive, spotted cDNA arrays. An Invention Disclosure has been filed by the Collaborating Investigator with the Fred Hutchinson Cancer Research Center (F.H.C.R.C.), Seattle, WA for this exciting new technology. The detection of SNPs on DNA microarrays has heretofore only been possible using allele specific hybridization on oligonucleotide arrays (GeneChips) produced by Affymetrix. Limited access to these GeneChips and their prohibitive costs will make it difficult to apply this technology to large numbers of DNA samples. A major goal of the proposed study is to develop and demonstrate the feasibility of the new SNP detection methodology for high throughput detection of polymorphisms at multiple loci using the high-density cDNA arrays produced by the novel arrayer to be developed in this grant. These highly reproducible, high-density arrays are also expected to provide improved sensitivity and specificity in the analysis of mRNA expression. Experiments will be performed to demonstrate the advantages of the proposed technology over existing pin based mechanical arrayers for gene expression analysis. PROPOSED COMMERCIAL APPLICATIONS: Fueled by emerging technologies like bio-chips, micro-arrays and high throughput screening and the high costs of reagents in general, the bio- medical industry is going towards assays using smaller fluid sample volumes. The proposed research directly addresses this growing market by developing a general purpose automated laboratory pipetting instrument for these applications. By keeping development costs down, the new technology will make high quality, high density DNA arrays accessible to a much broader segment of the cancer research community.