Rapid Screening of Nucleic Acid Energetics

描述(申请人提供)：最近的进展，可能是最明显的人类基因组计划，提出了新的和令人信服的手段和目标，以利用核酸杂交的精致特异性。为了合理设计和优化杂交探针，用于高通量筛选、基于核酸的诊断和治疗以及许多新兴的生物技术应用，对核酸(和核酸类似物)复合体随温度变化的稳定性的精确了解是必要的。拟议项目的主要目标是开发一种高通量手段，通过测量和分析光学监测的、温度诱导的有序-无序转变来获得准确的核酸络合物的热力学数据。该装置的操作类似于pH计，操作员将加载一个核酸复杂样品，并在短暂的稳定时间后显示数值结果。该装置将在几秒钟内提供完整的热力学分析，能够准确地预测核酸复合体在任何温度下的稳定性。与公共卫生相关：拟议项目的主要目标是开发一种高通量手段，通过测量和分析光学监测的、温度诱导的有序-无序转变，获得准确的核酸复合体的热力学和热力学数据。这些信息对设计和排除探针以及更多的核酸基础研究是有用的。

DESCRIPTION (provided by applicant): Recent advances, perhaps most visible in the Human Genome Initiative, have presented new and compelling means and objectives to exploit the exquisite specificity of nucleic acid hybridization. Precise knowledge of the temperature dependent stability of nucleic acid (and nucleic acid analog) complexes is necessary for the rational design and optimization of hybridization probes for use in high throughput screening, nucleic acid based diagnostics and therapeutics, and numerous emerging biotechnology applications. The primary objective of the proposed project is to develop a high-throughput means of obtaining accurate thermal and thermodynamic data for nucleic acid complexes by measurement and analysis of optically-monitored, temperature-induced, order-disorder transitions. The proposed device will operate analogously to a pH meter, the operator will load a nucleic acid complex sample and after a short settling time a numerical result will be displayed. The device will provide, in a matter of seconds, a complete thermodynamic analysis capable of predicting accurately the stability of the nucleic acid complex at any temperature. PUBLIC HEALTH RELEVANCE: The primary objective of the proposed project is to develop a high-throughput means of obtaining accurate thermal and thermodynamic data for nucleic acid complexes by measurement and analysis of optically- monitored, temperature-induced, order-disorder transitions. This information is useful in designing and troubleshooting probes and in more fundamental studies of nucleic acids.