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用于精确一维凝胶分析的系统,包括高分辨率定量足迹分析。

System for accurate one-dimensional gel analysis including high-resolution quantitative footprinting.

作者信息

Smith J, Singh M

机构信息

Medical Research Council, Cambridge, England, UK.

出版信息

Biotechniques. 1996 Jun;20(6):1082-7. doi: 10.2144/96206bc01.

Abstract

The interactive menu-driven computer program, Geltrak, displays and analyzes a one-dimensional (1-D) electrophoretic gel after digitization by either densitometry or storage-phosphor technology. The program is designed to analyze nucleic acid or protein gels in a variety of ways and includes a specific footprinting option. Overall accuracy for the entire process of 1-D electrophoretic gel band quantitation by peak profile fitting has been investigated. A standard control gel with a predetermined component ratio and minimal background was devised and subjected to electrophoresis, followed by three methods of digitization. The resulting images were analyzed by Geltrak, and their integrated peak area ratios were compared with their initial measured values. Individual band profiles from the control gel were used to develop an improved fitting function to represent the peak profile shape exactly. This function can be used in conjunction with a sophisticated profile-fitting technique to accurately predict the peak shapes from gels with background noise or overlapping bands.

摘要

交互式菜单驱动的计算机程序Geltrak,在通过密度测定法或存储磷光技术进行数字化处理后,显示并分析一维(1-D)电泳凝胶。该程序旨在以多种方式分析核酸或蛋白质凝胶,并包括一个特定的足迹分析选项。通过峰形拟合对一维电泳凝胶条带定量的整个过程的总体准确性进行了研究。设计了一种具有预定成分比例和最小背景的标准对照凝胶,进行电泳,然后采用三种数字化方法。所得图像由Geltrak分析,并将其积分峰面积比与其初始测量值进行比较。使用对照凝胶的单个条带图谱来开发改进的拟合函数,以精确表示峰形。该函数可与复杂的图谱拟合技术结合使用,以准确预测存在背景噪声或条带重叠的凝胶的峰形。

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