Resolving power: a quantitative measure of electrophoretic resolution.
Resolving power: a quantitative measure of electrophoretic resolution.
复制标题
分辨率：电泳分辨率的定量测量。
DOI：
10.1006/abio.1993.1211
复制
发表时间：
1993
影响因子：
2.9
通讯作者：
Sutherland,JC
中科院分区：
文献类型：
--
作者：
Ribeiro,EA;Sutherland,JC
分辨力是电泳系统分离类似大小的DNA（和其他）分子的能力的定量量度。它是一个无量纲的量，因此便于比较操作非常不同的电泳系统的性能。分辨力可以根据由凝胶或印迹上一系列完全分辨的条带组成的实验数据确定为分子长度的函数;不需要紧密间隔的条带。我们讨论的因素，如在一个特定的频带中的DNA的质量和空间分辨率的系统用于图像的DNA的分布在凝胶或印迹，而不是电泳系统的固有部分，可能会影响所观察到的分辨率。我们推导出一个经验的全球分散函数，适用于所有样品的电泳固定时间后获得的凝胶图像和图像，每个物种到达位于一个固定的距离从起始井的检测器。我们使用这个分散函数表明，通过延长电泳的时间或距离在静态的，均匀的电场中产生的分辨率的改善渐近接近一个极限值，这是DNA的长度的函数。当作为分子长度的函数绘制时，该极限值定义了表征特定电泳系统的性能的固有极限的包络（例如，电场强度、凝胶类型和浓度、缓冲液、温度）。比较静态场琼脂糖凝胶电泳分离103 - 105 bp长度DNA分子的常规方法与其它电泳方法的分辨能力，表明可以取得显著的改进。
Resolving power is a quantitative measure of the ability of an electrophoretic system to separate DNA (and other) molecules of similar size. It is a dimensionless quantity, and hence facilitates comparison of the performance of electrophoretic systems that operate very differently. Resolving power can be determined as a function of molecular length from experimental data consisting of a series of completely resolved bands on a gel or blot; closely spaced bands are not required. We discuss factors such as the mass of DNA in a particular band and the spatial resolution of the system used to image the distribution of DNA on a gel or blot that, while not an intrinsic part of the electrophoretic system, may influence the observed resolving power. We derive an empirical global dispersion function that applies both to images of gels obtained after a fixed time of electrophoresis of all the samples and to images obtained as each species reaches a detector located at a fixed distance from the starting well. We use this dispersion function to show that the improvement in resolving power produced by extending the time or distance of electrophoresis in a static, uniform electric field asymptotically approaches a limiting value that is a function of the length of the DNA. When plotted as a function of molecular length, this limiting value defines an envelope that characterizes the intrinsic limits of performance of a particular electrophoretic system (e.g., electric field strength, gel type and concentration, buffer, temperature). Comparing the resolving power of static field agarose gel electrophoresis as routinely practiced for separating DNA molecules from 103to 105bp long with other electrophoretic schemes suggests that significant improvements should be achievable.