Galat A
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA.
Electrophoresis. 1989 Oct;10(10):659-67. doi: 10.1002/elps.1150101002.
Computer-aided quantitative analysis of densitometric spectra is presented. The densitometric spectra are decomposed into component bands using the Powell and Marquardt minimizers. Several different functions for the component bands are utilized. It is shown that the densitometric spectra can be decomposed into component bands with high accuracy only if the proper shapes of the bands are chosen. The method described was used for quantitative analysis of densitometric spectra of DNA cleaved by neocarzinostatin. The procedure is general and can be applied to analyses of autoradiographic films and to direct scans of electrophoretic gels. It is shown that densitometric spectra which show highly overlapped bands are well approximated by asymmetric Cauchy and Gauss functions. Well separated densitometric bands which have substantial asymmetry can be fitted to more sophisticated shapes formed by combinations of symmetric and asymmetric Cauchy and Gauss functions. High accuracy fitting to asymmetric densitometric curves may only be achieved using the cosine function introduced by Mignot and Rondot, J. Appl. Cryst. 1976, 9,460-465.
本文介绍了密度光谱的计算机辅助定量分析。使用Powell和Marquardt最小化算法将密度光谱分解为成分带。对成分带采用了几种不同的函数。结果表明,只有选择合适的带形状,密度光谱才能高精度地分解为成分带。所描述的方法用于新制癌菌素切割的DNA密度光谱的定量分析。该程序具有通用性,可应用于放射自显影片的分析以及电泳凝胶的直接扫描。结果表明,具有高度重叠带的密度光谱可以用不对称柯西函数和高斯函数很好地近似。具有明显不对称性的分离良好的密度带可以拟合为由对称和不对称柯西函数与高斯函数组合形成的更复杂形状。只有使用Mignot和Rondot在《应用晶体学杂志》1976年第9卷第460 - 465页中引入的余弦函数,才能实现对不对称密度曲线的高精度拟合。