DNA melting apparatus for characterizing the thermal dissociation of DNA and supramolecular complexes in solution
DNA melting apparatus for characterizing the thermal dissociation of DNA and supramolecular complexes in solution
批准号:
374699-2009
负责人:
GibbsDavis, Julianne
金额:
$3.06万
依托单位:
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
项目状态:
已结题
起止时间:
2008-01-01 至 2009-12-31
中文摘要
需要一种DNA熔融装置来提供监测溶液中非共价络合物的缔合和解离的手段。我们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双链的稳定性。为了确定我们的合成修饰的有效性,我们迫切需要一种方法来监控由此产生的稳定性。DNA复合体的温度可变测量是监测DNA稳定性的最常见和最简单的方法,所建议的仪器是专门为这项任务设计的。有了这台设备,我们将能够开发出一种DNA复制系统,可以用作一步靶扩增和检测策略。这种方法有可能极大地提高检测设备的灵敏度,目前检测设备在其检测序列中需要单独的酶扩增步骤。开发一种基于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具有功能的复制组件。此外,该设备还将用于表征由基于氢键、pi-pi堆积和静电元素的合成超分子络合物组成的非共价组件。使用DNA熔化装置来监测这些复合体的解离和缔合,将使我们能够确定是什么控制了它们的分子识别行为。这些见解最终将有助于理解在生物系统中重要的更复杂的分子识别过程。
英文摘要
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.
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依托单位:
海外基金