DNA melting apparatus for characterizing the thermal dissociation of DNA and supramolecular complexes in solution

用于表征溶液中 DNA 和超分子复合物热解离的 DNA 熔解装置

• 批准号: 374699-2009
• 负责人: GibbsDavis, Julianne
• 金额: $ 3.06万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2008
• 资助国家: 加拿大
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=411991
• 关键词: DNA; melting; apparatus; characterizing; thermal

A DNA melting apparatus is requested to provide means for monitoring the association and dissociation of noncovalent complexes in solution. One major focus of our research program involves synthetically modifying the structure of DNA to control the stability of DNA duplexes in a DNA replication cycle. To determine the efficacy of our synthetic modifications, we urgently require a method for monitoring the resulting stability. Temperature-variable measurements of DNA complexes are the most common and simple method for monitoring DNA stability, and the proposed instrumentation is designed specifically for this task. With this equipment, we will be able to develop a DNA replicating system that can be utilized as a one-step target amplification and detection strategy. Such an approach has the potential for greatly improving the sensitivity of detection devices, which currently require separate enzymatic amplification steps in their detection sequence. Developing a self-replicating system based on DNA will also shine light on the origins of life, and how molecules with recognition properties are able to organize into functional, replicating assemblies. Additionally, this equipment will be utilized to characterize noncovalent assemblies composed of synthetic supramolecular complexes based on hydrogen-bonding, pi-pi stacking and electrostatic elements. Using the DNA melting apparatus to monitor the dissociation and association of these complexes will allow us to determine what governs their molecular recognition behavior. These insights will ultimately be helpful in understanding more complex molecular recognition processes important in biological systems.

要求DNA解链装置提供用于监测溶液中非共价复合物的缔合和解离的手段。 我们的研究计划的一个主要重点是合成修饰DNA的结构，以控制DNA复制周期中DNA双链体的稳定性。 为了确定我们的合成修饰的功效，我们迫切需要一种用于监测所得稳定性的方法。 DNA复合物的温度可变测量是用于监测DNA稳定性的最常见和最简单的方法，并且所提出的仪器是专门为该任务而设计的。 有了这台设备，我们将能够开发出一种DNA复制系统，可用作一步靶扩增和检测策略。 这种方法具有极大地提高检测装置的灵敏度的潜力，所述检测装置目前在其检测序列中需要单独的酶促扩增步骤。 开发基于DNA的自我复制系统也将揭示生命的起源，以及具有识别特性的分子如何能够组织成功能性的复制组件。 此外，该设备将用于表征由基于氢键、π-π堆积和静电元素的合成超分子复合物组成的非共价组装体。 使用DNA熔解装置来监测这些复合物的解离和缔合将使我们能够确定是什么支配它们的分子识别行为。这些见解最终将有助于理解生物系统中重要的更复杂的分子识别过程。