Cross-platform comparison of nucleic acid hybridization: toward quantitative reference standards.
Cross-platform comparison of nucleic acid hybridization: toward quantitative reference standards.
复制标题
核酸杂交的跨平台比较：针对定量参考标准。
DOI：
10.1016/j.ab.2014.08.001
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发表时间：
2014
影响因子：
2.9
通讯作者：
Agris,PaulF
中科院分区：
文献类型：
--
作者：
Halvorsen,Ken;Agris,PaulF
测量生物分子之间的相互作用对于基础研究和应用研究以及药物开发都是至关重要的。尽管有越来越多的技术可用于测量相互作用的生物分子的各种动力学和热力学性质，但很难比较不同实验室和人员的技术，甚至使用相同技术的不同仪器的数据。在这里，我们基于互补的DNA和RNA寡核苷酸来评估相关的生物相互作用，这些寡核苷酸可以作为许多实验系统的参考标准。我们用等温滴定量热法、差示扫描量热法和紫外-可见光(UV-Vis)监测变性/复性来测量双链形成的热力学。这些标准可用于验证结果，比较来自不同技术的数据，作为实验室课程的教学工具，或潜在地校准仪器。RNA和DNA标准具有许多吸引人的特点，包括低成本、高纯度、易于测量的浓度和最小的处理问题，使它们成为理想的参考材料。
Measuring interactions between biological molecules is vitally important to both basic and applied research as well as development of pharmaceuticals. Although a wide and growing range of techniques is available to measure various kinetic and thermodynamic properties of interacting biomolecules, it can be difficult to compare data across techniques of different laboratories and personnel or even across different instruments using the same technique. Here we evaluate relevant biological interactions based on complementary DNA and RNA oligonucleotides that could be used as reference standards for many experimental systems. We measured thermodynamics of duplex formation using isothermal titration calorimetry, differential scanning calorimetry, and ultraviolet–visible (UV–vis) monitored denaturation/renaturation. These standards can be used to validate results, compare data from disparate techniques, act as a teaching tool for laboratory classes, or potentially to calibrate instruments. The RNA and DNA standards have many attractive features, including low cost, high purity, easily measurable concentrations, and minimal handling concerns, making them ideal for use as a reference material.