HIGH SENSITIVITY, ULTRA-LOW BACKGROUND DNA CHIPS

开发高功能的DNA芯片需要整合各种技术和专业知识。理想的DNA芯片必须易于使用，允许高密度的靶DNA附着，显示极高的信噪比，在各种存储和打印条件下都是稳定的，并且具有成本效益。生产这样的芯片不仅需要将DNA附着在玻璃表面的能力，还需要创造出与这种芯片一起使用的理想荧光探针的技术和能力。该提案详细说明了如何使用这些独立但密切相关的技术(DNA芯片形成和荧光探针合成)来创建DNA芯片和检测系统，以实现最大的杂交灵敏度。在第一阶段，将测试各种不同的化合物是否有能力防止荧光探针DNA非特异性附着到改性玻璃表面。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高密度和稳定地附着在玻璃表面。在第二阶段，第一阶段开发的低背景芯片将被优化，用于使用各种高荧光探针(标记IT技术)进行杂交。使用这些探针，将阐明杂交和检测能力的关键参数的系统优化。拟议的商业应用：用于微阵列应用的一系列高效、易于使用的DNA芯片的开发和优化将对研究和诊断市场具有巨大的商业价值。这项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).