HIGH SENSITIVITY, ULTRA-LOW BACKGROUND DNA CHIPS

高功能DNA芯片的开发需要多种技术和专业知识的整合。理想的DNA芯片必须易于使用，允许高密度的目标DNA附着，显示极高的信噪比，在各种存储和打印条件下保持稳定，并且具有成本效益。生产这种芯片不仅需要将DNA附着在玻璃表面的能力，还需要技术和能力来创造用于该芯片的理想荧光探针。该提案详细介绍了这些独立但密切相关的技术（DNA芯片形成和荧光探针合成）将如何用于创建DNA芯片和检测系统，从而实现最大的杂交灵敏度。在第一阶段，各种不同的化合物将被测试其防止荧光探针DNA非特异性附着到修饰玻璃表面的能力。一旦确定，将制备DNA芯片（包含最具保护性的化合物），以促进目标DNA在玻璃表面的高密度和稳定附着。在第二阶段，在第一阶段开发的低背景芯片将使用各种高荧光探针（Label IT技术）进行杂交优化。利用这些探针，系统优化杂交的关键参数和检测能力将得到阐明。建议的商业应用：开发和优化一系列用于微阵列应用的高效，易于使用的DNA芯片将对研究和诊断市场具有巨大的商业价值。该技术的成功开发将产生两条高度优化的产品线，用于DNA芯片市场（即低背景DNA芯片和用于芯片应用的核酸标记试剂）。

The development of highly functional DNA chips requires the integration of a variety of technologies and expertise. The ideal DNA chip must be easy to use, allow high density target DNA attachment, display extremely high signal to noise ratios, be stable under various storage and printing conditions, and be cost effective. Production of such a chip requires not only the ability to attach DNA to a glass surface, but also the technology and capability to create the ideal fluorescent probes for use with this chip. This proposal details how these separate, but closely related, technologies (DNA chip formation and fluorescent probe synthesis) will be used to create a DNA chip and detection system that allows maximum hybridization sensitivity. During phase I, a variety of different compounds will be tested for their ability to prevent non-specific attachment of fluorescent probe DNA to a modified glass surface. Once identified, DNA chips (incorporating the most protective compounds) will be prepared that facilitate both high density and stable attachment of target DNA to the glass surface. During phase II, the low background chips developed during phase I will be optimized for hybridization using a variety of highly fluorescent probes (Label IT technology). Using these probes, the systematic optimization of critical parameters for hybridization and detection capabilities will be elucidated. PROPOSED COMMERCIAL APPLICATION: The development and optimization of a series of highly efficient, easy to use DNA chips for use in microarray applications will be of great commercial value for both the research and diagnostic markets. Successful development of this technology will result in two highly optimized product lines for use in the DNA chip market (i.e. low background DNA chips and nucleic acid labeling reagents for chip applications).